Uses of il-13 antagonists for treating atopic dermatitis

ABSTRACT

Uses of IL-13 antagonists for treating atopic dermatitis are provided. Also provided are methods of treating atopic dermatitis and methods of reducing the severity of atopic dermatitis by administering IL-13 antagonists.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of International Application No.PCT/US2017/052891 having an International Filing Date of Sep. 22, 2017,which claims the benefit of provisional U.S. Application No. 62/539,037,filed Jul. 31, 2017, provisional U.S. Application No. 62/530,683, filedJul. 10, 2017, provisional U.S. Application No. 62/527,204, filed Jun.30, 2017, and provisional U.S. Application No. 62/398,713, filed Sep.23, 2016 all of which are hereby incorporated by reference in theirentirety.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted via EFS-Web and is hereby incorporated by reference in itsentirety. Said ASCII copy, created on Mar. 12, 2019, is namedP33854-US-4 Sequence Listing.TXT and is 12,554 bytes in size.

FIELD

Uses of IL-13 antagonists for treating atopic dermatitis are provided.Also provided are methods of treating atopic dermatitis and methods ofreducing the severity of atopic dermatitis by administering IL-13antagonists.

BACKGROUND

Atopic dermatitis (AD) is a chronic relapsing and remitting inflammatoryskin disorder affecting all age groups. Clinically, AD is characterizedby xerosis, erythematous crusting rash, lichenification, an impairedskin barrier, and intense pruritus (Bieber T., N Engl J Med2008;358:1483-94).

Patients with AD have a high disease burden, and their quality of life(QoL) is significantly impacted. In one study, AD was shown to have agreater negative effect on patient mental health than diabetes andhypertension (Zuberbier T, et al., J Allergy Clin Immunol2006;118:226-32). Patients with moderate to severe AD have a higherprevalence of social dysfunction and sleep impairment, which is directlyrelated to severity of disease (Williams H, et al., J Allergy ClinImmunol 2008;121:947-54.e15). Depression, anxiety, and socialdysfunction not only affect patients with AD but also their caregivers(Zuberbier T, et al., J Allergy Clin Immunol 2006;118:226-32). Comparedwith psoriasis, another common and debilitating skin disease, ADpatients have lower role-physical, vitality, social functioning,role-emotional, and mental health subscale scores (Kiebert G, et al.,Int J Dermatol 2002;41:151-8).

Interleukin (IL)-13 is considered a key mediator of T-helper type 2(Th2) inflammation and elevated levels of IL-13 have been associated notonly with atopic dermatitis but with numerous other diseases including,but not limited to, asthma, inflammatory bowel disease, idiopathicpulmonary fibrosis (IPF), and chronic obstructive pulmonary disease(COPD) (Oh C K, et al., Eur Respir Rev 19:46-54 (2010); Fahy J V, etal., Nat Rev Immunol 15:57-65 [2015]). IL-13 is produced by many celltypes, including Th2 cells, basophils, eosinophils, and mast cells, aswell as airway epithelial cells and Type 2 innate lymphoid cells. IL-13binds to a heterodimeric receptor, IL-4Rα/IL-13Rα1 that is shared withIL-4 and activates the STAT-6 signaling pathway (Hershey G K, J AllergyClin Immunol 111(4):677-90 [2003]). Because Th2 inflammation involvesthe activity of several cell types in addition to Th2 cells, includingType 2 innate lymphoid cells (ILC2s), “Th2 inflammation” has morerecently been referred to in the scientific literature as “Type 2inflammation.” In addition to Th2 cells, ILC2s have been identified asimportant sources of cytokines such as IL-5 and IL-13. Accordingly,cytokines such as IL-13 and IL-5 that have been previously identified asTh2 cytokines are now also referred to as Type 2 cytokines in thescientific literature. Likewise, the disease states associated with suchcytokines, including atopic dermatitis, are now also referred to as Type2-driven diseases or Type 2-associated diseases. See, e.g., Noonan etal., J. Allergy Clin Immunol., 132(3): 567-574 (2013); Hanania et al.,Thorax 70(8): 748-56 (2015); and Cai et al., Bioanalysis 8(4): 323-332(2016).

Eosinophilic inflammation is associated with a variety of illnesses,both allergic and non-allergic (Gonlugur (2006) Immunol. Invest.35(1):29-45). Inflammation is a restorative response of living tissuesto injury. A characteristic of inflammatory reactions is theaccumulation of leukocytes in injured tissue due to certain chemicalsproduced in the tissue itself. Eosinophil leukocytes accumulate in awide variety of conditions such as allergic disorders, helminthicinfections, and neoplastic diseases (Kudlacz et al., (2002) Inflammation26: 111-119). Eosinophil leukocytes, a component of the immune system,are defensive elements of mucosal surfaces. They respond not only toantigens but to parasites, chemicals, and trauma.

Tissue eosinophilia occurs in skin diseases such as eczema, pemphigus,acute urticaria, and toxic epidermal necrolysis as well as in atopicdermatitis (Rzany et al., Br. J. Dermatol. 135: 6-11 (1996)).Eosinophils accumulate in the tissue and empty granule proteins inIgE-mediated allergic skin reactions (Nielsen et al., Ann. AllergyAsthma Immunol., 85: 489-494 (2001)). Eosinophils combined with mastcells are likely to cause joint inflammation (Miossec, J. Clin.Rheumatol. 3: 81-83 (1997)). Eosinophilic inflammation sometimesaccompanies joint trauma. Synovial fluid eosinophilia can be associatedwith diseases such as rheumatoid arthritis, parasitic disease,hypereosinophilic syndrome, Lyme disease, and allergic processes, aswell as hemarthrosis and arthrography (Atanes et al., Scand. J.Rheumatol., 25: 183-185 (1996)). Eosinophilic inflammation can affectbones as well (Yetiser et al., Int. J. Pediatr. Otorhinolaryngol., 62:169-173 (2002)). Examples of eosinophilic muscle disease includeeosinophilic perimyositis, eosinophilic polymyositis, and focaleosinophilic myositis (Lakhanpal et al., Semin. Arthritis Rheum., 17:331-231 (1988)). Eosinophilic inflammations affecting skeletal musclesmay be associated with parasite infections or drugs or features of somesystemic disorders of hypereosinophilia (e.g., idiopathichypereosinophilic syndrome and eosinophilia-myalgia syndrome.Eosinophils participate in the inflammatory response to epitopesrecognized by autoimmune antibodies (Engineer et al., Cytokine, 13:32-38 (2001)). Connective tissue diseases may lead to neutrophilic,eosinophilic, or lymphocytic vascular inflammations (Chen et al., J. Am.Acad. Dermatol., 35: 173-182 (1996)). Tissue and peripheral bloodeosinophilia can occur in active rheumatismal diseases. Elevation ofserum ECP levels in ankylosing spondylitis, a kind of connective tissuedisease, suggests that eosinophils are also involved in the underlyingprocess (Feltelius et al., Ann. Rheum. Dis., 46: 403-407 (1987)).Wegener's granulomatosis can rarely present with pulmonary nodules,pleural effusion, and peripheral blood eosinophilia (Krupsky et al.,Chest, 104: 1290-1292 (1993)).

Peripheral blood eosinophilia of at least 400/mm3 can occur in 7% ofcases of systemic sclerosis, 31% of cases of localized scleroderma, and61% of cases of eosinophilic fasciitis (Falanga, et al., J. Am. Acad.Dermatol., 17: 648-656 (1987)). Scleroderma yields an inflammatoryprocess closely resembling Meissner's and Auerbach's plexuses andconsists of mast cells and eosinophil leukocytes in the gastrointestinalsystem. Eosinophil-derived neurotoxins can contribute togastrointestinal motor dysfunction, as occurs in scleroderma(DeSchryver-Kecskemeti, et al. Arch. Pathol. Lab Med., 113: 394-398(1989)).

Eosinophils can accompany localized (Varga, et al., Curr. Opin.Rheumatol., 9: 562-570 (1997)) or systemic (Bouros et al., Am. J.Respir. Crit. Care Med., 165: 1581-1586 (2002)) connective tissueproliferation. They can incite fibrosis by inhibiting proteoglycandegradation in fibroblasts (Hernnas et al., Eur. J. Cell Biol., 59:352-363 (1992)), and fibroblasts mediate eosinophil survival bysecreting GM-CSF (Vancheri et al., Am. J. Respir. Cell Mol. Biol., 1:289-214 (1989)). Eosinophils can be found in nasal (Bacherct et al., J.allergy Clin. Immunol., 107: 607-614 (2001)), bronchial (Arguelles, etal., Arch. Intern. Med., 143: 570-571 (1983)), and gastrointestinalpolyp tissues (Assarian, et al., Hum. Pathol., 16: 311-312 (1985)).Likewise, eosinophils can be localized in inflammatory pseudotumors(myofibroblastic tumor). Eosinophils often accompany inflammatorypseudotumors in the orbital region, in which case the condition canmimic angioedema or allergic rhinoconjunctivitis (Li et al., Ann.Allergy, 69: 101-105 (1992)).

Eosinophilic inflammation can be found in tissue trauma (e.g., as aresult of surgery or injury). Eosinophilic inflammation can also beassociated with cardiovascular illnesses (e.g., eosinophilicmyocarditis, eosinophilic coronary arteritis, ischemic heart disease,acute myocardial infarction, cardiac rupture). Necrotic inflammatoryprocesses can also involve eosinophilic inflammation (polymyositis,coronary artery dissection, necrotizing lesions of neuro-Behcet'sdisease, dementia, cerebral infarction).

Several lines of evidence suggest that IL-13 is a key pathogeneticcomponent in atopic dermatitis (AD). Increased expression of IL-13 hasconsistently been reported in AD skin (Hamid Q, et al., J Allergy ClinImmunol 98:225-31 [1996]; Jeong C W, et al., Clin Exp Allergy 33:1717-24[2003]; Tazawa T, et al., Arch Dermatol Res 295:459-64 [2004]; Neis M M,et al., J Allergy Clin Immunol 118:930-7 [2006]; Suárez-Fariñas M, etal., J Allergy Clin Immunol 132:361-70 [2013]; Choy D F, et al., JAllergy Clin Immunol 130:1335-43 [2012]) and some reports suggest arelationship between IL-13 expression and the severity of disease (LaGrutta S, et al., Allergy 60:391-5 [2005]). Increased IL-13 has alsobeen reported in the serum of AD patients (Novak N, et al., J InvestDermatol 2002;119:870-5; International Patent Application No.PCT/US2016/022481 [publication no. WO2016149276]), and several studieshave reported an increase in IL-13-expressing T cells in the blood of ADpatients (Akdis M, et al., J Immunol 1997;159:4611-9; Aleksza M, et al.,Br J Dermatol 2002;147:1135-41; La Grutta S, et al., Allergy2005;60:391-5). IL-13 and its receptors have therefore becometherapeutic targets for the treatment of various Type 2inflammation-associated diseases including asthma, IPF and AD (Corren J,et al., N Eng J Med 365:1088-98 [2011]; Scheerens H, et al., Clin ExpAllergy 44:38-46 [2014]; Beck L A, et al., N Eng J Med 371:130-9 [2014];Thaci D, et al., Lancet 2016; 387:40-52). Additional publications thatdiscuss IL-13 in atopic dermatitis or lebrikizumab include He J Q, etal., Genes Immun 2003;4:385-89; Kim B E, et al. Clin Immunology2008;126, 332-7; Bhogal R K & Bona C A Int Rev Immunol 2008;27:472-96;Kim S T, et al. J Gene Med 2009;11:26-37; Bieber T, et al. J AllergyClin Immunol 2014;133:AB404; Thaci D, et al. J Allergy Clin Immunol2014;133:AB192; 2. Ultsch M, et al. J Mol Biol 2013; 425:1330-1339.

In addition, several clinical studies investigating agents with broadlyacting anti-inflammatory activity have demonstrated a reduction in IL-13expression that was associated with improved clinical disease. Forexample, nineteen adult patients with moderate to severe AD treated for12 weeks with cyclosporin A showed a reduction in skin expression ofIL-13 (Khattri S, et al., J Allergy Clin Immunol 2014;133(6):1626-34),ten pediatric patients treated with cyclosporin A micro-emulsion showeda reduction in blood IL-13 expressing CD3⁺ T cells (Bunikowski R, etal., Pediatr Allergy Immunol 2001;12:216-23), and twelve adults withmoderate to severe AD treated with narrow-band ultraviolet B showed asignificant decrease in IL-13 skin expression (Tintle S, et al., JAllergy Clin Immunol 2011;128:583-93.e1-4).

A number of IL-13 antagonists have been described and clinically testedin various Type 2 inflammation-associated diseases including asthma,COPD, and IPF. These include IMA-026, IMA-638 (also referred to as,anrukinzumab, INN No. 910649-32-0; QAX-576); tralokinumab (also referredto as CAT-354, CAS No. 1044515-88-9); and AER-001, ABT-308 (alsoreferred to as humanized 13C5.5 antibody). In addition, certainIL-4receptor alpha antagonists have been developed and these areantagonists of both IL-13 and IL-4. Examples of IL-4receptor alphaantagonists include AMG-317, AIR-645, dupilumab, which has been testedclinically in atopic dermatitis as well as asthma (see, e.g., Beck L A,et al., N Eng J Med 371:130-9 [2014]) and AER-001, IL4/IL-13 trap.Another IL-13 antagonist is lebrikizumab. Lebrikizumab is a humanizedmonoclonal immunoglobulin (Ig) G4 antibody (huIgG4) with a mutation inthe hinge region for increased stability. Lebrikizumab bindsspecifically to soluble human IL-13 with high affinity and neutralizesits functional activities with high potency. Lebrikizumab inhibits IL-13signaling through the IL-4Ralpha/IL-13Ralpha1 receptor. It blocks thebinding of IL-13 to IL-4Ralpha, but does not block the binding of IL-13to IL-13Ralpha1 or IL-13Ralpha2. Lebrikizumab has been described invarious publications and tested in several asthma studies (see, e.g.,Corren et al. (2011) N Engl J Med 365: 1088-98; Scheerens et al. (2012)Am J Respir Crit Care Med 185: A3960; Jia et al. (2012) J Allergy ClinImmunol 130: 647-654 e10; Hanania et al., Thorax 2015;70:748-756;Hanania et al., Lancet Respir Med 2016, available atdx(dot)doi(dot)org(slash)S2213-2600(16)30265-X, published online Sep. 5,2016; WO 2012/083132).

The therapeutic approach to AD primarily consists of trigger avoidance,skin hydration with bathing and use of emollients and anti-inflammatorytherapies consisting predominantly of topical corticosteroids (TCS). Inmany patients, treatment with TCS provides some measure of symptomaticrelief but does not adequately control their disease. In addition, TCSuse is associated with many comorbidities and limitations including highpatient burden. Review of literature (including PubMed, the Nankervissystematic review, the Global Resource for Eczema Trials [GREAT]database) of randomized, controlled, blinded clinical trials of systemicimmunosuppressant treatments and oral glucocorticoids for treatment ofmoderate-to-severe AD highlights that the use of conventional systemicimmunosuppressant treatments is often limited by significant sideeffects; additionally, their use is mainly off-label. These findingsindicate a substantial unmet need and the importance of developing drugstargeting the underlying AD specific pathways, such as the recentlyapproved anti-IL-4Rα antibody dupilumab. The promise for biologicaltherapy in refractory AD is to provide more effective therapy thatreduces the need for systemic immunosuppressive therapy and ultimatelyreduces the need for intensive TCS therapy. While dupilumab is the firstand only biologic approved for the treatment of adults withmoderate-to-severe AD, it is administered as an initial dose of 600 mg(two 300 mg subcutaneous injections), followed by 300 mg every otherweek (Simpson et al., N Engl J Med. 2016; 375(24): 2335-2348).

In those patients who have persistent moderate-to-severe disease notresponding adequately to TCS, recent guidelines outline a number ofstep-up therapeutic options (Ring J, et al., J Eur Acad DermatolVenereol 2012;26:1176-93; Schneider L, et. al., J Allergy Clin Immunol2013;131:295-9. el-27). The step-up options include topical calcineurininhibitors (TCIs), phototherapy, and immunosuppressive agents such asoral corticosteroids, cyclosporine, azathioprine, methotrexate, andmycophenolate. Amongst these, only cyclosporine is approved fortreatment of moderate to severe AD (nationally licensed in many Europeancountries, but not in the U.S.), and its use is limited to patients aged16 years and over (for a maximum of 8 weeks [NEORAL®]). Thoughcyclosporine is the most extensively studied systemic agent,interpretation of the trial results and applicability to clinicalpractice is limited by the trial designs (Schmitt J, et al., J Eur AcadDermatol Venereol 2007;21:606-619). Studies of other immunosuppressantagents such azathioprine and methotrexate consist mostly of casereports, with few reported randomized controlled trials (Haeck I M, etal., J Am Acad Dermatol 2011;64:1074-84; Schram M E, et al., J AllergyClin Immunol 2011;128:353-9). Prescribing practice demonstratessignificant country-specific variation, highlighted by a recent surveyof European dermatologists' which reported no clearly favored agent: 43%used cyclosporine, 31% oral corticosteroids, and 22% azathioprine(Proudfoot L E, et al., Br J Dermatol 2013;169:901-9).

Although the step-up therapies, including systemic immunosuppressants,used to treat patients with moderate to severe AD show evidence ofmodest to good efficacy, poor tolerability due to side effects limittheir prolonged use. Even in cases where cyclosporine has demonstratedsubstantial efficacy, approximately 50% of patients relapse within 2weeks, and 80% relapse within 6 weeks after cessation of therapy (Amor KT, et al., J Am Acad Dermatol 2010;63:925-46). The continued use ofthese agents despite harmful side effects and limitations indicates ahigh unmet medical need for safer and more effective therapies.

In attempts to find novel therapies for moderate to severe AD with anacceptable benefit-risk profile, a number of biologic agents thatspecifically target inflammatory cells and mediators have been tested(Taïeb A, et al., J Dtsch Dermatol Ges 2012;10:174-8; Guttman-Yassky E,et al., Expert Opin Biol Ther 2013;13:549-61). However, several of thesestudies and case reports were performed in only a small number ofpatients (as few as one or two) and have shown inconsistent efficacyand/or safety signals.

Accordingly, there is a need for new therapeutic treatments andtreatment regimens that will reduce the severity of atopic dermatitisdisease symptoms and maximize efficacy. In addition, there is a need fornew therapeutic treatments and treatment regimens that provide animproved safety profile with limited toxicity compared to existingtreatments or provide more tolerability or convenience for patientsthereby improving patient compliance with usage of therapeutic agentsand adherence to treatment regimens.

The invention described herein meets certain of the above-describedneeds and provides other benefits.

All references cited herein, including patent applications andpublications, are incorporated by reference in their entirety for anypurpose.

SUMMARY

The invention is based, at least in part, on the surprising andunexpected discovery that an anti-IL-13 antagonist monoclonal antibody,lebrikizumab, provides therapeutic benefit when administered to atopicdermatitis patients using the dosing regimens provided herein, includingpatients using topical corticosteroids concomitantly, as assessed byseveral efficacy outcome measures. Accordingly, provided herein are usesof IL-13 antagonists, including anti-IL-13 antibodies such aslebrikizumab, in treating atopic dermatitis and methods of treatingatopic dermatitis with IL-13 antagonists, including anti-IL-13antibodies such as lebrikizumab.

Accordingly, in one aspect, methods of treating atopic dermatitis in apatient comprising administering to the patient a pharmaceuticalcomposition comprising a therapeutically effective amount of an IL-13antagonist, wherein the pharmaceutical composition reduces diseaseseverity in the patient and wherein disease severity is assessed by anAtopic Dermatitis Disease Severity Outcome Measure (ADDSOM) areprovided. In some embodiments, the atopic dermatitis is moderate tosevere as determined by Rajka/Langeland criteria score and wherein theRajka/Langeland criteria score is determined to be between 4.5 and 9. Insome embodiments, the method further comprises administration of one ormore topical corticosteroids. In some embodiments, the one or moretopical corticosteroids is administered before administration of theIL-13 antagonist, at the same time as administration of the IL-13antagonist, or after administration of the IL-13 antagonist. In someembodiments, the one or more topical corticosteroids is selected fromtriamcinolone acetonide, hydrocortisone, and a combination oftriamcinolone acetonide and hydrocortisone. In some embodiments, thepatient is aged 12 years and over. In some embodiments, the patient isinadequately controlled on topical corticosteroids. In some embodiments,the IL-13 antagonist is a monoclonal anti-IL-13 antibody. In someembodiments, the anti-IL-13 antibody is an antibody comprising a VHcomprising HVR-H1, HVR-H2, and HVR-H3, wherein the respective VH HVRshave the amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 6, and SEQ IDNO: 7, and comprising a VL comprising HVR-L1, HVR-L2, and HVR-L3,wherein the respective VL HVRs have the amino acid sequence of SEQ IDNO: 8, SEQ ID NO: 9, and SEQ ID NO: 10. In certain embodiments, theanti-IL-13 antibody is an IgG1, IgG2, IgG3, or IgG4. In certainembodiments, the anti-IL-13 antibody is a human, humanized, or chimericantibody. In some embodiments, the anti-IL-13 antibody is an antibodycomprising a VH comprising a sequence selected from SEQ ID NO: 1 and SEQID NO: 3, and comprising a VL comprising a sequence selected from SEQ IDNO: 2 and SEQ ID NO: 4. In certain embodiments, the anti-IL-13 antibodyis a full-length antibody or a fragment thereof that binds human IL-13.In some embodiments, the anti-IL-13 antibody is IgG4. In one embodiment,the anti-IL-13 antibody is lebrikizumab. In one embodiment, theanti-IL-13 antibody comprises a heavy chain having the amino acidsequence of SEQ ID NO: 11 and a light chain having the amino acidsequence of SEQ ID NO: 12. In some embodiments, the IL-13 antagonist isadministered to the patient using a subcutaneous administration device.In certain such embodiments, the subcutaneous administration device isselected from a prefilled syringe, disposable pen injection device,microneedle device, microinfuser device, needle-free injection device,and autoinjector device. In one embodiment, the IL-13 antagonist islebrikizumab and lebrikizumab is administered to the patient using aprefilled syringe. In one embodiment, the IL-13 antagonist islebrikizumab and lebrikizumab is administered to the patient using anautoinjector device.

In another aspect, the pharmaceutical composition for use in treatingatopic dermatitis comprises 125 mg or 250 mg or 500 mg or about 125 mgor about 250 mg or about 500 mg of an anti-IL-13 antibody. In someembodiments, the pharmaceutical composition comprises between 110 mg and140 mg of an anti-IL-13 antibody or between 120 mg and 130 mg of ananti-IL-13 antibody. In some embodiments, the pharmaceutical compositioncomprises between 225 mg and 275 mg of an anti-IL-13 antibody or between240 mg and 260 mg of an anti-IL-13 antibody. In some embodiments, thepharmaceutical composition comprises between 450 mg and 550 mg of ananti-IL-13 antibody or between 475 mg and 525 mg of an anti-IL-13antibody or between 490 mg and 510 mg of an anti-IL-13 antibody. In someembodiments, the pharmaceutical composition comprises 125 mg, or about125 mg, or between 110 mg and 140 mg, or between 120 mg and 130 mg, ofthe anti-IL-13 antibody and the composition is administeredsubcutaneously once every four weeks. In some embodiments, thepharmaceutical composition is administered for a period of 12 weeks or aperiod of 20 weeks or a period of 24 weeks. In some embodiments, thepharmaceutical composition comprises 250 mg, or about 250 mg, or between225 mg and 275 mg, or between 240 mg and 260 mg of the anti-IL-13antibody and the composition is administered subcutaneously once everyfour weeks or once every eight weeks. In some embodiments, thepharmaceutical composition is administered for a period of 24 weeks ormore or for 24 weeks. In some embodiments, the atopic dermatitis ismoderate to severe as determined by Rajka/Langeland criteria score andwherein the Rajka/Langeland criteria score is determined to be between4.5 and 9. In some embodiments, the IL-13 antagonist is a monoclonalanti-IL-13 antibody. In some embodiments, the anti-IL-13 antibody is anantibody comprising a VH comprising HVR-H1, HVR-H2, and HVR-H3, whereinthe respective VH HVRs have the amino acid sequence of SEQ ID NO: 5, SEQID NO: 6, and SEQ ID NO: 7, and comprising a VL comprising HVR-L1,HVR-L2, and HVR-L3, wherein the respective VL HVRs have the amino acidsequence of SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10. In certainembodiments, the anti-IL-13 antibody is an IgG1, IgG2, IgG3, or IgG4. Incertain embodiments, the anti-IL-13 antibody is a human, humanized, orchimeric antibody. In some embodiments, the anti-IL-13 antibody is anantibody comprising a VH comprising a sequence selected from SEQ ID NO:1 and SEQ ID NO: 3, and comprising a VL comprising a sequence selectedfrom SEQ ID NO: 2 and SEQ ID NO: 4. In certain embodiments, theanti-IL-13 antibody is a full-length antibody or a fragment thereof thatbinds human IL-13. In some embodiments, the anti-IL-13 antibody is IgG4.In one embodiment, the anti-IL-13 antibody is lebrikizumab. In oneembodiment, the anti-IL-13 antibody comprises a heavy chain having theamino acid sequence of SEQ ID NO: 11 and a light chain having the aminoacid sequence of SEQ ID NO: 12. In some embodiments, the IL-13antagonist is administered to the patient using a subcutaneousadministration device. In certain such embodiments, the subcutaneousadministration device is selected from a prefilled syringe, disposablepen injection device, microneedle device, microinfuser device,needle-free injection device, and autoinjector device. In oneembodiment, the IL-13 antagonist is lebrikizumab and lebrikizumab isadministered to the patient using a prefilled syringe. In oneembodiment, the IL-13 antagonist is lebrikizumab and lebrikizumab isadministered to the patient using an autoinjector device. In someembodiments, the method further comprises administration of one or moretopical corticosteroids. In some embodiments, the one or more topicalcorticosteroids is administered before administration of the IL-13antagonist, at the same time as administration of the IL-13 antagonist,or after administration of the IL-13 antagonist. In some embodiments,the one or more topical corticosteroids is selected from triamcinoloneacetonide, hydrocortisone, and a combination of triamcinolone acetonideand hydrocortisone. In some embodiments, the patient is aged 12 yearsand over. In some embodiments, the patient is inadequately controlled ontopical corticosteroids.

In yet another aspect, methods of treating atopic dermatitis in apatient comprising administering to the patient a pharmaceuticalcomposition comprising a therapeutically effective amount of an IL-13antagonist, wherein the pharmaceutical composition reduces diseaseseverity in the patient and wherein disease severity is assessed by anAtopic Dermatitis Disease Severity Outcome Measure (ADDSOM) areprovided, and wherein the Atopic Dermatitis Disease Severity OutcomeMeasure is Eczema Area and Severity Index (EAST), or Severity Scoring ofAtopic Dermatitis (SCORAD), or Investigator Global Assessment (IGA), ora Patient Reported Outcome (PRO). In some embodiments, the atopicdermatitis is moderate to severe as determined by Rajka/Langelandcriteria score and wherein the Rajka/Langeland criteria score isdetermined to be between 4.5 and 9. In some embodiments, the methodfurther comprises administration of one or more topical corticosteroids.In some embodiments, the one or more topical corticosteroids isadministered before administration of the IL-13 antagonist, at the sametime as administration of the IL-13 antagonist, or after administrationof the IL-13 antagonist. In some embodiments, the one or more topicalcorticosteroids is selected from triamcinolone acetonide,hydrocortisone, and a combination of triamcinolone acetonide andhydrocortisone. In some embodiments, the patient is aged 12 years andover. In some embodiments, the patient is inadequately controlled ontopical corticosteroids. In some embodiments, the ADDSOM is EASI and thepharmaceutical composition reduces the EASI by 50% or 75% or 90%compared to the EASI determined prior to administration of a first doseof the pharmaceutical composition. In some embodiments, the EASI isdetermined 12 weeks after administration of the first dose or 20 weeksafter administration of the first dose or 24 weeks after administrationof the first dose. In some embodiments, the ADDSOM is SCORAD and thepharmaceutical composition reduces the SCORAD by 50% compared to theSCORAD determined prior to administration of a first dose of thepharmaceutical composition. In some embodiments, the ADDSOM is SCORADand the pharmaceutical composition reduces the SCORAD by 75% compared tothe SCORAD determined prior to administration of a first dose of thepharmaceutical composition. In some embodiments, the SCORAD isdetermined 12 weeks after administration of the first dose. In someembodiments, the ADDSOM is IGA and the pharmaceutical compositionreduces the IGA to zero or one. In some embodiments, the IGA isdetermined 12 weeks after administration of a first dose of thepharmaceutical composition. In some embodiments, the ADDSOM is PRO andthe PRO is pruritus visual analogue scale (VAS), sleep loss VAS, orAtopic Dermatitis Impact Questionnaire (ADIQ) score. In someembodiments, the PRO is determined 12 weeks after administration of afirst dose of the pharmaceutical composition. In some embodiments, thePRO is pruritus VAS and the pharmaceutical composition reduces thepruritus VAS by 40% to 55%. In some embodiments, the PRO is sleep lossVAS and the pharmaceutical composition reduces the sleep loss VAS by 53%to 61%. In some embodiments, the PRO is ADIQ and the pharmaceuticalcomposition reduces the ADIQ score by 54% to 65%. In some embodiments,the IL-13 antagonist is a monoclonal anti-IL-13 antibody. In someembodiments, the anti-IL-13 antibody is an antibody comprising a VHcomprising HVR-H1, HVR-H2, and HVR-H3, wherein the respective VH HVRshave the amino acid sequence of SEQ ID NO: 5, SEQ ID NO: 6, and SEQ IDNO: 7, and comprising a VL comprising HVR-L1, HVR-L2, and HVR-L3,wherein the respective VL HVRs have the amino acid sequence of SEQ IDNO: 8, SEQ ID NO: 9, and SEQ ID NO: 10. In certain embodiments, theanti-IL-13 antibody is an IgG1, IgG2, IgG3, or IgG4. In certainembodiments, the anti-IL-13 antibody is a human, humanized, or chimericantibody. In some embodiments, the anti-IL-13 antibody is an antibodycomprising a VH comprising a sequence selected from SEQ ID NO: 1 and SEQID NO: 3, and comprising a VL comprising a sequence selected from SEQ IDNO: 2 and SEQ ID NO: 4. In certain embodiments, the anti-IL-13 antibodyis a full-length antibody or a fragment thereof that binds human IL-13.In some embodiments, the anti-IL-13 antibody is IgG4. In one embodiment,the anti-IL-13 antibody is lebrikizumab. In one embodiment, theanti-IL-13 antibody comprises a heavy chain having the amino acidsequence of SEQ ID NO: 11 and a light chain having the amino acidsequence of SEQ ID NO: 12. In some embodiments, the IL-13 antagonist isadministered to the patient using a subcutaneous administration device.In certain such embodiments, the subcutaneous administration device isselected from a prefilled syringe, disposable pen injection device,microneedle device, microinfuser device, needle-free injection device,and autoinjector device. In one embodiment, the IL-13 antagonist islebrikizumab and lebrikizumab is administered to the patient using aprefilled syringe. In one embodiment, the IL-13 antagonist islebrikizumab and lebrikizumab is administered to the patient using anautoinjector device.

In still yet another aspect, methods comprising administering to apatient a therapeutically effective amount of an IL-13 antagonist,wherein the therapeutically effective amount is selected from 125 mg and250 mg and wherein the IL-13 antagonist is administered subcutaneouslyonce every four weeks are provided. In some embodiments, thetherapeutically effective amount is about 125 mg or between 110 mg and140 mg of an anti-IL-13 antibody or between 120 mg and 130 mg of ananti-IL-13 antibody. In some embodiments, the therapeutically effectiveamount is about 250 mg or between 225 mg and 275 mg of an anti-IL-13antibody or between 240 mg and 260 mg of an anti-IL-13 antibody.

In some embodiments, the IL-13 antagonist is a monoclonal anti-IL-13antibody. In some embodiments, the anti-IL-13 antibody is an antibodycomprising a VH comprising HVR-H1, HVR-H2, and HVR-H3, wherein therespective VH HVRs have the amino acid sequence of SEQ ID NO: 5, SEQ IDNO: 6, and SEQ ID NO: 7, and comprising a VL comprising HVR-L1, HVR-L2,and HVR-L3, wherein the respective VL HVRs have the amino acid sequenceof SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10. In certainembodiments, the anti-IL-13 antibody is an IgG1, IgG2, IgG3, or IgG4. Incertain embodiments, the anti-IL-13 antibody is a human, humanized, orchimeric antibody. In some embodiments, the anti-IL-13 antibody is anantibody comprising a VH comprising a sequence selected from SEQ ID NO:1 and SEQ ID NO: 3, and comprising a VL comprising a sequence selectedfrom SEQ ID NO: 2 and SEQ ID NO: 4. In certain embodiments, theanti-IL-13 antibody is a full-length antibody or a fragment thereof thatbinds human IL-13. In some embodiments, the anti-IL-13 antibody is IgG4.In one embodiment, the anti-IL-13 antibody is lebrikizumab. In oneembodiment, the anti-IL-13 antibody comprises a heavy chain having theamino acid sequence of SEQ ID NO: 11 and a light chain having the aminoacid sequence of SEQ ID NO: 12. In some embodiments, the IL-13antagonist is administered to the patient using a subcutaneousadministration device. In certain such embodiments, the subcutaneousadministration device is selected from a prefilled syringe, disposablepen injection device, microneedle device, microinfuser device,needle-free injection device, and autoinjector device. In oneembodiment, the IL-13 antagonist is lebrikizumab and lebrikizumab isadministered to the patient using a prefilled syringe. In oneembodiment, the IL-13 antagonist is lebrikizumab and lebrikizumab isadministered to the patient using an autoinjector device. In someembodiments, the atopic dermatitis is moderate to severe as determinedby Rajka/Langeland criteria score and wherein the Rajka/Langelandcriteria score is determined to be between 4.5 and 9. In someembodiments, the method further comprises administration of one or moretopical corticosteroids. In some embodiments, the one or more topicalcorticosteroids is administered before administration of the IL-13antagonist, at the same time as administration of the IL-13 antagonist,or after administration of the IL-13 antagonist. In some embodiments,the one or more topical corticosteroids is selected from triamcinoloneacetonide, hydrocortisone, and a combination of triamcinolone acetonideand hydrocortisone. In some embodiments, the patient is aged 12 yearsand over. In some embodiments, the patient is inadequately controlled ontopical corticosteroids. In some embodiments, the therapeuticallyeffective amount reduces disease severity in the patient and diseaseseverity is assessed by an Atopic Dermatitis Disease Severity OutcomeMeasure (ADDSOM). In some embodiments, the ADDSOM is Eczema Area andSeverity Index (EAST), or Severity Scoring of Atopic Dermatitis(SCORAD), or Investigator Global Assessment (IGA), or a Patient ReportedOutcome (PRO). In some embodiments, the ADDSOM is EASI and thetherapeutically effective amount reduces the EASI by 50% or 75% or 90%compared to the EASI determined prior to administration of a first doseof the IL-13 antagonist. In some embodiments, the EASI is determined 12weeks after administration of the first dose or 20 weeks afteradministration of the first dose or 24 weeks after administration of thefirst dose. In some embodiments, the ADDSOM is SCORAD and thetherapeutically effective amount reduces the SCORAD by 50% compared tothe SCORAD determined prior to administration of a first dose of theIL-13 antagonist. In some embodiments, the ADDSOM is SCORAD and thetherapeutically effective amount reduces the SCORAD by 75% compared tothe SCORAD determined prior to administration of a first dose of theIL-13 antagonist. In some embodiments, the SCORAD is determined 12 weeksafter administration of the first dose. In some embodiments, the ADDSOMis IGA and the therapeutically effective amount reduces the IGA to zeroor one. In some embodiments, the IGA is determined 12 weeks afteradministration of a first dose of the IL-13 antagonist. In someembodiments, the ADDSOM is PRO and the PRO is pruritus visual analoguescale (VAS), sleep loss VAS, or Atopic Dermatitis Impact Questionnaire(ADIQ) score. In some embodiments, the PRO is determined 12 weeks afteradministration of a first dose of the IL-13 antagonist. In someembodiments, the PRO is pruritus VAS and the therapeutically effectiveamount reduces the pruritus VAS by 40% to 55%. In some embodiments, thePRO is sleep loss VAS and the therapeutically effective amount reducesthe sleep loss VAS by 53% to 61%. In some embodiments, the PRO is ADIQand the therapeutically effective amount reduces the ADIQ score by 54%to 65%.

In another aspect, methods of treating atopic dermatitis in a patient byadministering to the patient a therapeutically effective amount of anIL-13 antagonist, where the administration comprises administration ofat least one loading dose and administration of at least one subsequentmaintenance dose, and where each of the at least one loading dose andeach of the at least one maintenance dose is administered subcutaneouslyat a flat dose are provided. In some embodiments, the atopic dermatitisis moderate to severe as determined by Rajka/Langeland criteria scoreand wherein the Rajka/Langeland criteria score is determined to bebetween 4.5 and 9. In some embodiments, the method further comprisesadministration of one or more topical corticosteroids. In someembodiments, the one or more topical corticosteroids is administeredbefore administration of the IL-13 antagonist, at the same time asadministration of the IL-13 antagonist, or after administration of theIL-13 antagonist. In some embodiments, the one or more topicalcorticosteroids is selected from triamcinolone acetonide,hydrocortisone, and a combination of triamcinolone acetonide andhydrocortisone. In some embodiments, the patient is aged 12 years andover. In some embodiments, the patient is inadequately controlled ontopical corticosteroids. In some embodiments, the IL-13 antagonist is amonoclonal anti-IL-13 antibody. In some embodiments, the anti-IL-13antibody is an antibody comprising a VH comprising HVR-H1, HVR-H2, andHVR-H3, wherein the respective VH HVRs have the amino acid sequence ofSEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7, and comprising a VLcomprising HVR-L1, HVR-L2, and HVR-L3, wherein the respective VL HVRshave the amino acid sequence of SEQ ID NO: 8, SEQ ID NO: 9, and SEQ IDNO: 10. In certain embodiments, the anti-IL-13 antibody is an IgG1,IgG2, IgG3, or IgG4. In certain embodiments, the anti-IL-13 antibody isa human, humanized, or chimeric antibody. In some embodiments, theanti-IL-13 antibody is an antibody comprising a VH comprising a sequenceselected from SEQ ID NO: 1 and SEQ ID NO: 3, and comprising a VLcomprising a sequence selected from SEQ ID NO: 2 and SEQ ID NO: 4. Incertain embodiments, the anti-IL-13 antibody is a full-length antibodyor a fragment thereof that binds human IL-13. In some embodiments, theanti-IL-13 antibody is IgG4. In one embodiment, the anti-IL-13 antibodyis lebrikizumab. In one embodiment, the anti-IL-13 antibody comprises aheavy chain having the amino acid sequence of SEQ ID NO: 11 and a lightchain having the amino acid sequence of SEQ ID NO: 12. In someembodiments, the loading dose is 250 mg or 500 mg and the maintenancedose is 125 mg. In some embodiments, the loading dose is 250 mg and themaintenance dose is 125 mg. In some embodiments, the loading dose is 500mg and the maintenance dose is 125 mg. In some embodiments, themaintenance dose is administered four weeks after administration of theloading dose and the maintenance dose is administered and once everyfour weeks thereafter for the duration of treatment. In someembodiments, the loading dose is 250 mg and the maintenance dose is 125mg, where the maintenance dose is administered four weeks after theloading dose and once every four weeks thereafter for the duration oftreatment. In some embodiments, the loading dose is 250 mg and theloading dose is administered followed by a second loading doseadministration 15 days later and the maintenance dose is 125 mg. In someembodiments, the maintenance dose is administered two weeks afteradministration of the second loading dose and the maintenance dose isadministered once every four weeks thereafter for the duration oftreatment. In some embodiments, the loading dose is 250 mg and theloading dose is administered followed by a second loading doseadministration 29 days later and the maintenance dose is 125 mg. In someembodiments, the maintenance dose is administered four weeks afteradministration of the second loading dose and the maintenance dose isadministered once every four weeks thereafter for the duration oftreatment. In some embodiments, the loading dose is 500 mg and themaintenance dose is 250 mg. In some embodiments, the maintenance dose isadministered four weeks after administration of the loading dose and themaintenance dose is administered once every four weeks thereafter forthe duration of treatment. In some embodiments, the maintenance dose isadministered four weeks after administration of the loading dose and themaintenance dose is administered once every eight weeks thereafter forthe duration of treatment. In some embodiments, the loading dose is 500mg and the maintenance dose is 250 mg, where the maintenance dose isadministered four weeks after the loading dose and once every four weeksthereafter for the duration of treatment. In some embodiments, theduration of treatment is 24 weeks or more. In some embodiments, theduration of treatment is 24 weeks. In some embodiments, thetherapeutically effective amount reduces disease severity in the patientand disease severity is assessed by an Atopic Dermatitis DiseaseSeverity Outcome Measure (ADDSOM). In some embodiments, the ADDSOM isEczema Area and Severity Index (EASI), or Severity Scoring of AtopicDermatitis (SCORAD), or Investigator Global Assessment (IGA), or aPatient Reported Outcome (PRO). In some embodiments, the ADDSOM is EASIand the therapeutically effective amount reduces the EASI by 50% or 75%or 90% compared to the EASI determined prior to administration of afirst dose of the IL-13 antagonist. In some embodiments, the EASI isdetermined 12 weeks after administration of the first dose or 20 weeksafter administration of the first dose or 24 weeks after administrationof the first dose. In some embodiments, the ADDSOM is SCORAD and thetherapeutically effective amount reduces the SCORAD by 50% compared tothe SCORAD determined prior to administration of a first dose of theIL-13 antagonist. In some embodiments, the ADDSOM is SCORAD and thetherapeutically effective amount reduces the SCORAD by 75% compared tothe SCORAD determined prior to administration of a first dose of theIL-13 antagonist. In some embodiments, the SCORAD is determined 12 weeksafter administration of the first dose. In some embodiments, the ADDSOMis IGA and the therapeutically effective amount reduces the IGA to zeroor one. In some embodiments, the IGA is determined 12 weeks afteradministration of a first dose of the IL-13 antagonist. In someembodiments, the ADDSOM is PRO and the PRO is pruritus visual analoguescale (VAS), sleep loss VAS, or Atopic Dermatitis Impact Questionnaire(ADIQ) score. In some embodiments, the PRO is determined 12 weeks afteradministration of a first dose of the IL-13 antagonist. In someembodiments, the PRO is pruritus VAS and the therapeutically effectiveamount reduces the pruritus VAS by 40% to 55%. In some embodiments, thePRO is sleep loss VAS and the therapeutically effective amount reducesthe sleep loss VAS by 53% to 61%. In some embodiments, the PRO is ADIQand the therapeutically effective amount reduces the ADIQ score by 54%to 65%. In some embodiments, the IL-13 antagonist is administered to thepatient using a subcutaneous administration device. In certain suchembodiments, the subcutaneous administration device is selected from aprefilled syringe, disposable pen injection device, microneedle device,microinfuser device, needle-free injection device, and autoinjectordevice. In one embodiment, the IL-13 antagonist is lebrikizumab andlebrikizumab is administered to the patient using a prefilled syringe.In one embodiment, the IL-13 antagonist is lebrikizumab and lebrikizumabis administered to the patient using an autoinjector device.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the Study I Schema as described in Example 2. Abbreviationsare as follows: D=day; Pbo=placebo; SC=subcutaneous; TCS=topicalcorticosteroid; Wk=week.

FIG. 2A shows the proportion of patients achieving EASI-50 over 12 weeksas described in Example 2. Dashed line with open circles, lebrikizumab250 mg single dose plus topical corticosteroids (TCS) twice per day(BID); dotted line with closed triangles, lebrikizumab 125 mg singledose plus TCS twice per day; solid line with open squares, lebrikizumab125 mg once every 4 weeks (Q4W) plus TCS twice per day; solid line withplus signs, placebo plus TCS twice per day.

FIG. 2B shows the proportion of patients achieving SCORAD-50 over 12weeks as described in Example 2. Dashed line with open circles,lebrikizumab 250 mg single dose plus topical corticosteroids (TCS) twiceper day (BID); dotted line with closed triangles, lebrikizumab 125 mgsingle dose plus TCS twice per day; solid line with open squares,lebrikizumab 125 mg once every 4 weeks (Q4W) plus TCS twice per day;solid line with plus signs, placebo plus TCS twice per day.

FIG. 2C shows the proportion of patients achieving IGA 0/1 over 12 weeksas described in Example 2. Dashed line with open circles, lebrikizumab250 mg single dose plus topical corticosteroids (TCS) twice per day(BID); dotted line with closed triangles, lebrikizumab 125 mg singledose plus TCS twice per day; solid line with open squares, lebrikizumab125 mg once every 4 weeks (Q4W) plus TCS twice per day; solid line withplus signs, placebo plus TCS twice per day.

FIGS. 3A-3C show the proportion of patients achieving EASI-50 (FIG. 3A),EASI-75 (FIG. 3B) and EASI-90 (FIG. 3C) over 20 weeks in the modifiedintention-to-treat population as described in Example 2. Dashed linewith open circles, lebrikizumab 250 mg single dose plus topicalcorticosteroids (TCS) twice per day (BID); dotted line with closedtriangles, lebrikizumab 125 mg single dose plus TCS twice per day; solidline with open squares, lebrikizumab 125 mg once every 4 weeks (Q4W)plus TCS twice per day; solid line with plus signs, placebo plus TCStwice per day.

FIG. 4 shows the percent change from baseline in EASI over 12 weeks inthe modified intention-to-treat population as described in Example 2.Dashed line with open circles, lebrikizumab 250 mg single dose plustopical corticosteroids (TCS) twice per day (BID); dotted line withclosed triangles, lebrikizumab 125 mg single dose plus TCS twice perday; solid line with open squares, lebrikizumab 125 mg once every 4weeks (Q4W) plus TCS twice per day; solid line with plus signs, placeboplus TCS twice per day. Data points shown are adjusted mean withstandard error bars. The asterisk (*) indicates that the placebocorrected change is 17.4%, P=0.025.

FIG. 5 shows the atopic dermatitis longitudinal PK-PD model oflebrikizumab as described in Example 3. In FIG. 5, R(t) is the EASIscore; K_(in0) is the baseline disease progression rate constant; k_(in)is the disease progression rate constant with lebrikizumab and includingthe placebo/TCS effect; E_(drug) is the lebrikizumab drug effect ondisease progression inhibition; E_(con) is the placebo/TCS effect(constant overtime); E_(max) is the maximum lebrikizumab drug effect ondisease progression inhibition; EC₅₀ is the lebrikizumab concentrationthat leads to 50% of E_(max); k_(out) is the tissue repair rateconstant; R(t=0) is the baseline EASI score; IIV is the inter-individualvariability; ω_(kout) is the inter-individual variability for k_(out);ω_(Econ) is the inter-individual variability for E_(con); ρ_(KoutXEcon)is the correlation between k_(out) and E_(con); and σ is the residualerror.

FIG. 6 shows the model-predicted median EASI-75 responses over time forGroup 1 (see Table 5) lebrikizumab dosing regimens as described inExample 3. The heavy solid line is the median simulated response forplacebo treatment (the improvement in EASI-75 in placebo treatmentrepresents the likely contribution of topical corticosteroids to theoverall efficacy). The medium solid line is the median simulatedresponse for the 125 mg lebrikizumab administered once every four weeks(Lebri 125 mg Q4W) dosing regimen. The dot-dash line is the mediansimulated response for a 250 mg loading dose given on Day 1 followed bya 125 mg maintenance dose of lebrikizumab administered once every fourweeks (Lebri 250 mg load, 125 mg Q4W) beginning at Week 4. The lightsolid line is the median simulated response for a 500 mg loading dosegiven at Day 1 followed by a 125 mg maintenance dose of lebrikizumabadministered once every four weeks (Lebri 500 mg load, 125 mg Q4W)beginning at Week 4. The heavy dashed line is the median simulatedresponse for a 250 mg dose of lebrikizumab administered on Days 1 and 29followed by a 125 mg maintenance dose of lebrikizumab administered onceevery four weeks (Lebri 250 mg days 1 & 29, 125 mg Q4W) starting at Week8. The light dashed line is the median simulated response for a 250 mgdose of lebrikizumab administered on Days 1 and 15 followed by a 125 mgmaintenance dose of lebrikizumab administered once every four weeks(Lebri 250 mg days 1 & 15, 125 mg Q4W) beginning at Week 4 (Day 29).Confidence intervals were removed from the plots for clarity.

FIG. 7 shows the model-predicted median EASI-75 responses over time forGroup 2 (see Table 5) lebrikizumab dosing regimens as described inExample 3. The heavy solid line is the median simulated response forplacebo treatment (the improvement in EASI-75 in placebo treatmentrepresents the likely contribution of topical corticosteroids to theoverall efficacy). The medium solid line is the median simulatedresponse for the 125 mg lebrikizumab administered once every four weeks(Lebri 125 mg Q4W) dosing regimen. The light dashed line is the mediansimulated response for the 250 mg lebrikizumab administered once everyfour weeks (Lebri 250 mg Q4W) dosing regimen. The heavy dashed line is a500 mg loading dose administered at Day 1 followed by a 250 mgmaintenance dose of lebrikizumab administered once every four weeksbeginning at week 4 (Lebri 500 mg load, 250 mg Q4W). Confidenceintervals were removed from the plots for clarity.

FIG. 8 shows the model-predicted median EASI-75 responses over time forGroup 3 (see Table 5) dosing regimens as described in Example 3. Theheavy solid line is the median simulated response for placebo treatment(the improvement in EASI-75 in placebo treatment represents the likelycontribution of topical corticosteroids to the overall efficacy). Themedium solid line is the median simulated response for the 125 mglebrikizumab administered once every four weeks (Lebri 125 mg Q4W)dosing regimen. The light dashed line is the median simulated responsefor 250 mg lebrikizumab administered once every eight weeks (Lebri 250Q8W). The heavy dashed line is the median simulated response for a 500mg loading dose followed by a maintenance dose of 250 mg lebrikizumabadministered once every eight weeks beginning on week 4 (Lebri 500 mgload, 250 mg Q8W). Confidence intervals were removed from the plots forclarity.

FIG. 9 shows the model-predicted median EASI-75 responses over time forGroup 4 (see Table 5) lebrikizumab dosing regimens as described inExample 3. The heavy solid line is the median simulated response forplacebo treatment (the improvement in EASI-75 in placebo treatmentrepresents the likely contribution of topical corticosteroids to theoverall efficacy). The medium solid line is the median simulatedresponse for the 125 mg lebrikizumab administered once every four weeks(Lebri 125 mg Q4W) dosing regimen. The heavy dashed line is the mediansimulated response for 37.5 mg lebrikizumab administered once every fourweeks dosing regimen (Lebri 37.5 mg Q4W). The light dashed line is themedian simulated response for a 125 mg loading dose followed by amaintenance dose of 37.5 mg lebrikizumab administered once every fourweeks beginning on week 4 (Lebri 125 mg load, 37.5 mg Q4W). Confidenceintervals were removed from the plots for clarity.

FIG. 10 shows the Study II Schema as described in Example 4.Abbreviations are as follows: D =day; EASI =Eczema Area and SeverityIndex; q4wk =every 4 weeks; TCS =topical corticosteroid; Wk =week.

FIGS. 11A-11D show the proportion of patients achieving EASI-50 (FIG.11A), EASI-75 (FIG. 11B), IGA 0/1 (FIG. 11C), and SCORAD-50 (FIG. 11D)at Week 12 as described in Example 2 and the change in the proportion ofpatients achieving EASI-50, EASI-75, IGA 0/1, and SCORAD-50,respectively, in each arm of the study compared to placebo is indicatedby the dotted line and arrows adjacent to each bar in each of FIGS.11A-11D. Abbreviations are as follows: EASI=Eczema Area Severity Index;IGA=Investigator Global Assessment; Q4W=every 4 weeks; SCORAD=SCORingAtopic Dermatitis; SD=single dose. Spotted bar=lebrikizumab 125 mgsingle dose plus topical corticosteroids (TCS) twice per day (BID);hatched bar=lebrikizumab 250 mg single dose plus TCS twice per day;stippled bar=lebrikizumab 125 mg once every 4 weeks (Q4W) plus TCS twiceper day; open bar=placebo plus TCS twice per day.

FIGS. 12A-12D show the change from baseline in pruritus VAS (FIG. 12A),ADIQ (FIG. 12B), sleep loss VAS (FIG. 12C), and DLQI (FIG. 12D) overtime as described in Example 2. Dashed line with open circles,lebrikizumab 250 mg single dose plus topical corticosteroids (TCS) twiceper day (BID); dotted line with closed triangles, lebrikizumab 125 mgsingle dose plus TCS twice per day; solid line with open squares,lebrikizumab 125 mg once every 4 weeks (Q4W) plus TCS twice per day;solid line with plus signs, placebo plus TCS twice per day.Abbreviations are as follows: VAS=Visual Analog Score; ADIQ=AtopicDermatitis Impact Questionnaire; DLQI=Dermatology Life Quality Index.

DETAILED DESCRIPTION

All references cited herein, including patent applications andpublications, are incorporated by reference in their entirety for anypurpose.

Unless defined otherwise, technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Singleton et al., Dictionary ofMicrobiology and Molecular Biology 2nd ed., J. Wiley & Sons (New York,N.Y. 1994), and March, Advanced Organic Chemistry Reactions, Mechanismsand Structure 4th ed., John Wiley & Sons (New York, N.Y. 1992), provideone skilled in the art with a general guide to many of the terms used inthe present application.

Certain Definitions

For purposes of interpreting this specification, the followingdefinitions will apply and whenever appropriate, terms used in thesingular will also include the plural and vice versa. In the event thatany definition set forth below conflicts with any document incorporatedherein by reference, the definition set forth below shall control.

As used in this specification and the appended claims, the singularforms “a,” “an” and “the” include plural referents unless the contextclearly dictates otherwise. Thus, for example, reference to “a protein”or an “antibody” includes a plurality of proteins or antibodies,respectively; reference to “a cell” includes mixtures of cells, and thelike.

Ranges provided in the specification and appended claims include bothend points and all points between the end points. Thus, for example, arange of 2.0 to 3.0 includes 2.0, 3.0, and all points between 2.0 and3.0

The terms “marker” and “biomarker” are used interchangeably to refer toa molecule, including a gene, protein, carbohydrate structure, orglycolipid, metabolite, mRNA, miRNA, protein, DNA (cDNA or genomic DNA),DNA copy number, or an epigenetic change, e.g., increased, decreased, oraltered DNA methylation (e.g., cytosine methylation, or CpG methylation,non-CpG methylations); histone modification (e.g., (de)acetylation, (de)methylation,(de) phosphorylation, ubiquitination, SUMOylation,ADP-ribosylation); altered nucleosome positioning, the expression orpresence of which in or on a mammalian tissue or cell can be detected bystandard methods (or methods disclosed herein) and which may bepredictive, diagnostic and/or prognostic for a mammalian cell's ortissue's sensitivity to treatment regimes based on Type-2 inflammationpathway inhibition using, for example, a Type-2 inflammation pathwayinhibitor such as an anti-IL-13 antibody as described herein. Abiomarker may also be a biological or clinical attribute that can bemeasured in a biological sample obtained from a subject, such as forexample but not limited to, blood cell count.

The term “biological sample” includes, but is not limited to, blood,serum, plasma, peripheral blood mononuclear cells (PBMCs), sputum,tissue biopsies (e.g., lung samples), and nasal samples including nasalswabs or nasal polyps. The sample may be taken before treatment, duringtreatment or post-treatment.

The term “atopic dermatitis” or “AD” means a chronic relapsing andremitting inflammatory skin disorder characterized by intense pruritus(e.g., severe itch), xerosis (e.g., abnormally dry skin), erythematouscrusting rash, lichenification, an impaired skin barrier and by scalyand dry eczematous lesions. The term “atopic dermatitis” includes, butis not limited to, AD caused by or associated with epidermal barrierdysfunction, allergy (e.g., allergy to certain foods, pollen, mold, dustmite, animals, etc.), radiation exposure, and/or asthma. In many cases,chronic AD lesions include thickened plaques of skin, lichenificationand fibrous papules.

A therapeutic agent a provided herein includes an agent that can bind tothe target cytokine, interleukin (IL)-13, such as a polypeptide(s)(e.g., an antibody, an immunoadhesin or a peptibody), an aptamer or asmall molecule that can bind to a protein or a nucleic acid moleculethat can bind to a nucleic acid molecule encoding a target identifiedherein (i.e., siRNA).

“Anti-IL-13 binding agent” refers to an agent that binds to human IL-13.Such binding agents can include a small molecule, aptamer or apolypeptide. Such polypeptide can include, but is not limited to, apolypeptide(s) selected from the group consisting of an immunoadhesin,an antibody, a peptibody and a peptide. According to one embodiment, thebinding agent binds to a human IL-13 sequence with an affinity between 1uM-1 pM. Specific examples of anti-IL-13 binding agents can includeanti-IL-13 antibodies, soluble IL-13receptoralpha2 fused to a human Fc,soluble IL4receptoralpha fused to a human Fc, soluble IL-13receptoralpha fused to a human Fc. Exemplary anti-IL-13 antibodiesinclude, but are not limited to, IMA-026, IMA-638 (also referred to as,anrukinzumab, INN No. 910649-32-0; QAX-576), tralokinumab (also referredto as CAT-354, CAS No. 1044515-88-9); AER-001, ABT-308 (also referred toas humanized 13C5.5 antibody, and lebrikizumab. According to oneembodiment, the anti-IL-13 antibody comprises a VH comprising a sequenceselected from SEQ ID NOs: 1, 3, and 24, and a VL comprising a sequenceselected from SEQ ID NO: 2, 4, and 25. In one embodiment, the anti-IL-13antibody comprises HVRH1, HVRH2, HVRH3, HVRL1, HVRL2, and HVRL3, whereinthe respective HVRs have the amino acid sequence of SEQ ID NO.: 5, SEQID NO.: 6, SEQ ID NO.: 7, SEQ ID NO.: 8, SEQ ID NO.: 9, and SEQ ID NO.:10. In one embodiment, the anti-IL-13 antibody is lebrikizumab.According to one embodiment, the antibody is an IgG1 antibody. Accordingto another embodiment, the antibody is an IgG4 antibody. According toone embodiment, the IgG4 antibody comprises a S228P mutation in itsconstant domain. In one embodiment, the anti-IL-13 antibody comprises aQ1E mutation in its variable heavy chain region. In one embodiment, theanti-IL-13 antibody comprises a M4L mutation in its variable light chainregion.

The term “small molecule” refers to an organic molecule having amolecular weight between 50 Daltons to 2500 Daltons.

The term “antibody” is used in the broadest sense and specificallycovers, for example, monoclonal antibodies, polyclonal antibodies,antibodies with polyepitopic specificity, single chain antibodies,multi-specific antibodies, including bispecific antibodies, and antibodyfragments so long as they exhibit the desired antigen-binding activity.Such antibodies can be chimeric, humanized, human and synthetic.

The term “uncontrolled” or “uncontrollable” refers to the inadequacy ofa treatment regimen to minimize a symptom of a disease. As used herein,the term “uncontrolled” and “inadequately controlled” can be usedinterchangeably and are meant to refer to the same state. The controlstatus of a patient can be determined by the attending physician basedon a number of factors including the patient's clinical history,responsiveness to treatment and level of current treatment prescribed.

The term “therapeutic agent” refers to any agent that is used to treat adisease.

The term “corticosteroid sparing” or “CS” means the decrease infrequency and/or amount, or the elimination of, corticosteroid used totreat a disease in a patient taking corticosteroids for the treatment ofthe disease due to the administration of another therapeutic agent. A“CS agent” refers to a therapeutic agent that can cause CS in a patienttaking a corticosteroid.

The term “corticosteroid” includes, but is not limited to, topicalcorticosteroids. Exemplary topical corticosteroids include triamcinoloneacetonide, typically formulated at a concentration of 0.1% in a cream,and hydrocortisone, typically formulated at a concentration of 1% or2.5% in a cream. Certain topical corticosteroids are considered veryhigh potency such as, for example, betamethasone dipropionate,clobetasol propionate, diflorasone diacetate, fluocinonide, andhalobetasol propionate. Certain topical corticosteroids are consideredhigh potency such as, for example, amcinonide, desoximetasone,halcinonide, and triamcinolone acetonide. Certain topicalcorticosteroids are considered medium potency, such as, for example,betamethasone valerate, clocortolone pivalate, fluocinolone acetonide,flurandrenolide, fluocinonide, fluticasone propionate, hydrocortisonebutyrate, hydrocortisone valerate, mometasone furoate, andprednicarbate. Certain topical corticosteroids are considered lowpotency, such as, for example, alclometasone dipropionate, desonide, andhydrocortisone. “Inhalable corticosteroid” means a corticosteroid thatis suitable for delivery by inhalation. Exemplary inhalablecorticosteroids are fluticasone, beclomethasone dipropionate,budenoside, mometasone furoate, ciclesonide, flunisolide, triamcinoloneacetonide and any other corticosteroid currently available or becomingavailable in the future. Examples of corticosteroids that can be inhaledand are combined with a long-acting beta2-agonist include, but are notlimited to: budesonide/formoterol and fluticasone/salmeterol.

The term “loading dose” means a dose of a drug given at the beginning ofa course of treatment that is higher than the dose given subsequentlyand each dose given for the remainder of the treatment, which isreferred to as a “maintenance dose.” Typically, a loading dose isadministered once or twice. After administration of the loading dose orloading doses, a maintenance dose is administered and the maintenancedose is administered thereafter, typically at regular intervals, for theremainder of the course of treatment.

The term “flat dose” means that a single dose is used for all patientsregardless of weight or any patient-specific factors related to weightor body mass. For example, administration of a flat dose of 100 mg of anantibody means that every patient, regardless of weight, will receive adose of 100 mg. A flat dose is sometimes referred to as a fixed dose.

The term “weight-based dose” means a dose that is calculated based onthe patient's weight. Thus, the dose administered depends on thepatient's weight. For example, a dose of 1 mg/kg of an antibody meansthat a patient weighing 50 kg will receive a dose of 50 mg while apatient weighing 80 kg will receive a dose of 80 mg.

“Patient response” or “response” (and grammatical variations thereof) toa therapeutic agent can be assessed using any endpoint indicating abenefit to the patient, including, without limitation, (1) inhibition,to some extent, of disease progression, including slowing down andcomplete arrest; (2) reduction in the number of disease episodes and/orsymptoms; (3) reduction in lesional size; (4) inhibition (i.e.,reduction, slowing down or complete stopping) of immune or inflammatorycell infiltration into adjacent peripheral organs and/or tissues; (5)inhibition (i.e. reduction, slowing down or complete stopping) ofdisease spread; (6) decrease of auto-immune response, which may, butdoes not have to, result in the regression or ablation of the diseaselesion; (7) relief, to some extent, of one or more symptoms associatedwith the disorder; and/or (8) increase in the length of disease-freepresentation following treatment.

“A patient maintains responsiveness to a treatment” when the patient'sresponsiveness does not decrease with time during the course of atreatment. A patient is an “inadequate responder” when the patient'sresponsiveness decreases with time during the course of treatment. Forexample, an atopic dermatitis patient whose symptoms were controlled bytopical corticosteroids (TCS) at the beginning of treatment but whosesymptoms fail to be relieved by TCS administration at later times duringthe course of treatment is losing responsiveness to treatment and isconsidered an inadequate responder to TCS.

A “subcutaneous administration device” refers to a device which isadapted or designed to administer a drug, for example a therapeuticantibody, or pharmaceutical formulation by the subcutaneous route.Exemplary subcutaneous administration devices include, but are notlimited to, a syringe, including a pre-filled syringe, an injectiondevice, infusion pump, injector pen, needleless device, and patchdelivery system. A subcutaneous administration device administers acertain volume of the pharmaceutical formulation, for example about 1.0mL, about 1.25 mL, about 1.5 mL, about 1.75 mL, or about 2.0 mL.

“Affinity” refers to the strength of the sum total of noncovalentinteractions between a single binding site of a molecule (e.g., anantibody) and its binding partner (e.g., an antigen). Unless indicatedotherwise, as used herein, “binding affinity” refers to intrinsicbinding affinity which reflects a 1:1 interaction between members of abinding pair (e.g., antibody and antigen binding arm). The affinity of amolecule X for its partner Y can generally be represented by thedissociation constant (Kd). Affinity can be measured by common methodsknown in the art, including those described herein. Specificillustrative and exemplary embodiments for measuring binding affinityare described in the following.

An “affinity matured” antibody refers to an antibody with one or morealterations in one or more hypervariable regions (HVRs), compared to aparent antibody which does not possess such alterations, suchalterations resulting in an improvement in the affinity of the antibodyfor antigen.

The terms “anti-target antibody” and “an antibody that binds to target”refer to an antibody that is capable of binding the target withsufficient affinity such that the antibody is useful as a diagnosticand/or therapeutic agent in targeting the target. In one embodiment, theextent of binding of an anti-target antibody to an unrelated, non-targetprotein is less than about 10% of the binding of the antibody to targetas measured, e.g., by a radioimmunoassay (MA) or biacore assay. Incertain embodiments, an antibody that binds to a target has adissociation constant (Kd) of ≤1 M, ≤100 nM, ≤10 nM, ≤1 nM, ≤0.1 nM,≤0.01 nM, or ≤0.001 nM (e.g. 10-8 M or less, e.g. from 10-8 M to 10-13M, e.g., from 10-9 M to 10-13 M). In certain embodiments, an anti-targetantibody binds to an epitope of a target that is conserved amongdifferent species.

An “antibody fragment” refers to a molecule other than an intactantibody that comprises a portion of an intact antibody that binds theantigen to which the intact antibody binds. Examples of antibodyfragments include but are not limited to single chain Fv, Fab, Fab′,Fab′-SH, F(ab′)2; diabodies; linear antibodies; single-chain antibodymolecules (e.g. scFv); and multispecific antibodies formed from antibodyfragments.

An “antibody that binds to the same epitope” as a reference antibodyrefers to an antibody that blocks binding of the reference antibody toits antigen in a competition assay by 50% or more, and conversely, thereference antibody blocks binding of the antibody to its antigen in acompetition assay by 50% or more. Various methods for carrying outcompetition assays are well-known in the art.

An “acceptor human framework” for the purposes herein is a frameworkcomprising the amino acid sequence of a light chain variable domain (VL)framework or a heavy chain variable domain (VH) framework derived from ahuman immunoglobulin framework or a human consensus framework, asdefined below. An acceptor human framework “derived from” a humanimmunoglobulin framework or a human consensus framework may comprise thesame amino acid sequence thereof, or it may contain amino acid sequencechanges. In some embodiments, the number of amino acid changes are 10 orless, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less,3 or less, or 2 or less. In some embodiments, the VL acceptor humanframework is identical in sequence to the VL human immunoglobulinframework sequence or human consensus framework sequence.

The term “chimeric” antibody refers to an antibody in which a portion ofthe heavy and/or light chain is derived from a particular source orspecies, while the remainder of the heavy and/or light chain is derivedfrom a different source or species.

The “class” of an antibody refers to the type of constant domain orconstant region possessed by its heavy chain. There are five majorclasses of antibodies: IgA, IgD, IgE, IgG, and IgM, and several of thesemay be further divided into subclasses (isotypes), e.g., IgG1, IgG2,IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains thatcorrespond to the different classes of immunoglobulins are called α, δ,ϵ, γ, and μ, respectively.

“Effector functions” refer to those biological activities attributableto the Fc region of an antibody, which vary with the antibody isotype.Examples of antibody effector functions include: Clq binding andcomplement dependent cytotoxicity (CDC); Fc receptor binding;antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g. B cell receptor); and B cellactivation.

An “effective amount” of an agent, e.g., a pharmaceutical formulation,refers to an amount effective, at dosages and for periods of timenecessary, to achieve the desired therapeutic or prophylactic result.

The term “Fc region” herein is used to define a C-terminal region of animmunoglobulin heavy chain that contains at least a portion of theconstant region. The term includes native sequence Fc regions andvariant Fc regions. In one embodiment, a human IgG heavy chain Fc regionextends from Cys226, or from Pro230, to the carboxyl-terminus of theheavy chain. However, the C-terminal lysine (Lys447) of the Fc regionmay or may not be present. Unless otherwise specified herein, numberingof amino acid residues in the Fc region or constant region is accordingto the EU numbering system, also called the EU index, as described inKabat et al., Sequences of Proteins of Immunological Interest, 5th Ed.Public Health Service, National Institutes of Health, Bethesda, MD,1991.

“Framework” or “FR” refers to variable domain residues other thanhypervariable region (HVR) residues. The FR of a variable domaingenerally consists of four FR domains: FR1, FR2, FR3, and FR4.Accordingly, the HVR and FR sequences generally appear in the followingsequence in VH (or VL): FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4.

The terms “full length antibody,” “intact antibody,” and “wholeantibody” are used herein interchangeably to refer to an antibody havinga structure substantially similar to a native antibody structure orhaving heavy chains that contain an Fc region as defined herein.

The terms “host cell,” “host cell line,” and “host cell culture” areused interchangeably and refer to cells into which exogenous nucleicacid has been introduced, including the progeny of such cells. Hostcells include “transformants” and “transformed cells,” which include theprimary transformed cell and progeny derived therefrom without regard tothe number of passages. Progeny may not be completely identical innucleic acid content to a parent cell, but may contain mutations. Mutantprogeny that have the same function or biological activity as screenedor selected for in the originally transformed cell are included herein.

A “human antibody” is one which possesses an amino acid sequence whichcorresponds to that of an antibody produced by a human or a human cellor derived from a non-human source that utilizes human antibodyrepertoires or other human antibody-encoding sequences. This definitionof a human antibody specifically excludes a humanized antibodycomprising non-human antigen-binding residues.

A “human consensus framework” is a framework which represents the mostcommonly occurring amino acid residues in a selection of humanimmunoglobulin VL or VH framework sequences. Generally, the selection ofhuman immunoglobulin VL or VH sequences is from a subgroup of variabledomain sequences. Generally, the subgroup of sequences is a subgroup asin Kabat et al., Sequences of Proteins of Immunological Interest, FifthEdition, NIH Publication 91-3242, Bethesda MD (1991), vols. 1-3. In oneembodiment, for the VL, the subgroup is subgroup kappa I as in Kabat etal., supra. In one embodiment, for the VH, the subgroup is subgroup IIIas in Kabat et al., supra.

A “humanized” antibody refers to a chimeric antibody comprising aminoacid residues from non-human HVRs and amino acid residues from humanFRs. In certain embodiments, a humanized antibody will comprisesubstantially all of at least one, and typically two, variable domains,in which all or substantially all of the HVRs (e.g., CDRs) correspond tothose of a non-human antibody, and all or substantially all of the FRscorrespond to those of a human antibody. A humanized antibody optionallymay comprise at least a portion of an antibody constant region derivedfrom a human antibody. A “humanized form” of an antibody, e.g., anon-human antibody, refers to an antibody that has undergonehumanization.

The term “hypervariable region” or “HVR” refers to each of the regionsof an antibody variable domain which are hypervariable in sequenceand/or form structurally defined loops (“hypervariable loops”).Generally, native four-chain antibodies comprise six HVRs; three in theVH (H1, H2, H3), and three in the VL (L1, L2, L3). HVRs generallycomprise amino acid residues from the hypervariable loops and/or fromthe “complementarity determining regions” (CDRs), the latter typicallybeing of highest sequence variability and/or involved in antigenrecognition. An HVR region as used herein comprise any number ofresidues located within positions 24-36 (for HVRL1), 46-56 (for HVRL2),89-97 (for HVRL3), 26-35B (for HVRH1), 47-65 (for HVRH2), and 93-102(for HVRH3).

An “individual” or “patient” or “subject” is a mammal. Mammals include,but are not limited to, domesticated animals (e.g., cows, sheep, cats,dogs, and horses), primates (e.g., humans and non-human primates such asmonkeys), rabbits, and rodents (e.g., mice and rats). In certainembodiments, the individual or patient or subject is a human. In someembodiments, an “individual” or “patient” or “subject” herein is anysingle human subject eligible for treatment who is experiencing or hasexperienced one or more signs, symptoms, or other indicators of asthmaor a respiratory condition. Intended to be included as a subject are anysubjects involved in clinical research trials not showing any clinicalsign of disease, or subjects involved in epidemiological studies, orsubjects once used as controls. The subject may have been previouslytreated with an IL-13 antagonist or another drug, or not so treated. Thesubject may be naïve to an IL-13 antagonist when the treatment herein isstarted, i.e., the subject may not have been previously treated with,for example, an IL-13 antagonist at “baseline” (i.e., at a set point intime before the administration of a first dose of an IL-13 antagonist inthe treatment method herein, such as the day of screening the subjectbefore treatment is commenced). Such “naïve” subjects are generallyconsidered to be candidates for treatment with such drug(s).

A “pediatric” individual or patient or subject is a human from birth to18 years old (or 0 to 18 years old). In some embodiments, a pediatricindividual or patient or subject is from 2 to 6, 2 to 17, 6 to 11, 6 to18, 6 to 17, 8 to 17, 12 to 17, or 12 to 18 years old.

An “isolated” antibody is one which has been separated from a componentof its natural environment. In some embodiments, an antibody is purifiedto greater than 95% or 99% purity as determined by, for example,electrophoretic (e.g., SDS-PAGE, isoelectric focusing (IEF), capillaryelectrophoresis) or chromatographic (e.g., ion exchange or reverse phaseHPLC). For review of methods for assessment of antibody purity, see,e.g., Flatman et al., J. Chromatogr. B 848:79-87 (2007).

An “isolated” nucleic acid refers to a nucleic acid molecule that hasbeen separated from a component of its natural environment. An isolatednucleic acid includes a nucleic acid molecule contained in cells thatordinarily contain the nucleic acid molecule, but the nucleic acidmolecule is present extrachromosomally or at a chromosomal location thatis different from its natural chromosomal location.

“Isolated nucleic acid encoding an anti-target antibody” refers to oneor more nucleic acid molecules encoding antibody heavy and light chains(or fragments thereof), including such nucleic acid molecule(s) in asingle vector or separate vectors, and such nucleic acid molecule(s)present at one or more locations in a host cell.

The term “monoclonal antibody” refers to an antibody obtained from apopulation of substantially homogeneous antibodies, i.e., the individualantibodies comprising the population are identical and/or bind the sameepitope, except for possible variant antibodies, e.g., containingnaturally occurring mutations or arising during production of amonoclonal antibody preparation, such variants generally being presentin minor amounts. In contrast to polyclonal antibody preparations, whichtypically include different antibodies directed against differentdeterminants (epitopes), each monoclonal antibody of a monoclonalantibody preparation is directed against a single determinant on anantigen. Thus, the modifier “monoclonal” indicates the character of theantibody as being obtained from a substantially homogeneous populationof antibodies, and is not to be construed as requiring production of theantibody by any particular method. For example, the monoclonalantibodies to be used according to the methods provided herein may bemade by a variety of techniques, including but not limited to thehybridoma method, recombinant DNA methods, phage-display methods, andmethods utilizing transgenic animals containing all or part of the humanimmunoglobulin loci, such methods and other exemplary methods for makingmonoclonal antibodies being described herein.

A “naked antibody” refers to an antibody that is not conjugated to aheterologous moiety (e.g., a cytotoxic moiety) or radiolabel. The nakedantibody may be present in a pharmaceutical formulation.

“Native antibodies” refer to naturally occurring immunoglobulinmolecules with varying structures. For example, native IgG antibodiesare heterotetrameric glycoproteins of about 150,000 daltons, composed oftwo identical light chains and two identical heavy chains that aredisulfide-bonded. From N- to C-terminus, each heavy chain has a variableregion (VH), also called a variable heavy domain or a heavy chainvariable domain, followed by three constant domains (CH1, CH2, and CH3).Similarly, from N- to C-terminus, each light chain has a variable region(VL), also called a variable light domain or a light chain variabledomain, followed by a constant light (CL) domain. The light chain of anantibody may be assigned to one of two types, called kappa (κ) andlambda (λ), based on the amino acid sequence of its constant domain.

The term “package insert” is used to refer to instructions customarilyincluded in commercial packages of therapeutic products, that containinformation about the indications, usage, dosage, administration,combination therapy, contraindications and/or warnings concerning theuse of such therapeutic products. The term “package insert” is also usedto refer to instructions customarily included in commercial packages ofdiagnostic products that contain information about the intended use,test principle, preparation and handling of reagents, specimencollection and preparation, calibration of the assay and the assayprocedure, performance and precision data such as sensitivity andspecificity of the assay.

“Percent (%) amino acid sequence identity” with respect to a referencepolypeptide sequence is defined as the percentage of amino acid residuesin a candidate sequence that are identical with the amino acid residuesin the reference polypeptide sequence, after aligning the sequences andintroducing gaps, if necessary, to achieve the maximum percent sequenceidentity, and not considering any conservative substitutions as part ofthe sequence identity. Alignment for purposes of determining percentamino acid sequence identity can be achieved in various ways that arewithin the skill in the art, for instance, using publicly availablecomputer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR)software. Those skilled in the art can determine appropriate parametersfor aligning sequences, including any algorithms needed to achievemaximal alignment over the full length of the sequences being compared.For purposes herein, however, % amino acid sequence identity values aregenerated using the sequence comparison computer program ALIGN-2. TheALIGN-2 sequence comparison computer program was authored by Genentech,Inc., and the source code has been filed with user documentation in theU.S. Copyright Office, Washington D.C., 20559, where it is registeredunder U.S. Copyright Registration No. TXU510087. The ALIGN-2 program ispublicly available from Genentech, Inc., South San Francisco, Calif., ormay be compiled from the source code. The ALIGN-2 program should becompiled for use on a UNIX operating system, including digital UNIXV4.0D. All sequence comparison parameters are set by the ALIGN-2 programand do not vary.

In situations where ALIGN-2 is employed for amino acid sequencecomparisons, the % amino acid sequence identity of a given amino acidsequence A to, with, or against a given amino acid sequence B (which canalternatively be phrased as a given amino acid sequence A that has orcomprises a certain % amino acid sequence identity to, with, or againsta given amino acid sequence B) is calculated as follows:

100 times the fraction X/Y where X is the number of amino acid residuesscored as identical matches by the sequence alignment program ALIGN-2 inthat program's alignment of A and B, and where Y is the total number ofamino acid residues in B. It will be appreciated that where the lengthof amino acid sequence A is not equal to the length of amino acidsequence B, the % amino acid sequence identity of A to B will not equalthe % amino acid sequence identity of B to A. Unless specifically statedotherwise, all % amino acid sequence identity values used herein areobtained as described in the immediately preceding paragraph using theALIGN-2 computer program.

The term “pharmaceutical formulation” or “pharmaceutical composition”refers to a preparation which is in such form as to permit thebiological activity of an active ingredient contained therein to beeffective, and which contains no additional components which areunacceptably toxic to a subject to which the formulation would beadministered.

A “pharmaceutically acceptable carrier” refers to an ingredient in apharmaceutical formulation, other than an active ingredient, which isnontoxic to a subject. A pharmaceutically acceptable carrier includes,but is not limited to, a buffer, excipient, stabilizer, or preservative.

The term “target” refers to any native molecule from any vertebratesource, including mammals such as primates (e.g. humans) and rodents(e.g., mice and rats), unless otherwise indicated. The term encompasses“full-length,” unprocessed target as well as any form of target thatresults from processing in the cell. The term also encompasses naturallyoccurring variants of targets, e.g., splice variants or allelicvariants.

The term “treatment” (and grammatical variations thereof such as “treat”or “treating”) refers to clinical intervention in an attempt to alterthe natural course of the individual being treated, and can be performedeither for prophylaxis or during the course of clinical pathology.Desirable effects of treatment include, but are not limited to,preventing occurrence or recurrence of disease, alleviation of symptoms,diminishment of any direct or indirect pathological consequences of thedisease, preventing metastasis, decreasing the rate of diseaseprogression, amelioration or palliation of the disease state, andremission or improved prognosis. In some embodiments, antibodies areused to delay development of a disease or to slow the progression of adisease.

The term “variable region” or “variable domain” refers to the domain ofan antibody heavy or light chain that is involved in binding theantibody to antigen. The variable domains of the heavy chain and lightchain (VH and VL, respectively) of a native antibody generally havesimilar structures, with each domain comprising four conserved frameworkregions (FRs) and three hypervariable regions (HVRs). (See, e.g., Kindtet al. Kuby Immunology, 6th ed., W. H. Freeman and Co., page 91 (2007).)A single VH or VL domain may be sufficient to confer antigen-bindingspecificity. Furthermore, antibodies that bind a particular antigen maybe isolated using a VH or VL domain from an antibody that binds theantigen to screen a library of complementary VL or VH domains,respectively. See, e.g., Portolano et al., J. Immunol. 150:880-887(1993); Clarkson et al., Nature 352:624-628 (1991).

The term “vector” refers to a nucleic acid molecule capable ofpropagating another nucleic acid to which it is linked. The termincludes the vector as a self-replicating nucleic acid structure as wellas the vector incorporated into the genome of a host cell into which ithas been introduced. Certain vectors are capable of directing theexpression of nucleic acids to which they are operatively linked. Suchvectors are referred to herein as “expression vectors.”

The term “flare” in a patient diagnosed with atopic dermatitis means ameasurable increase in extent or severity of lesions over a period of atleast 3 days, under continued treatment and corresponding with aclinically significant increase in disease severity (as assessed by thetreating physician and/or by the patient) necessitating an escalation intherapy (see, e.g., Darsow et al., J Eur Acad Dermatol Venereol 2010;24:317-28).

Atopic dermatitis can be diagnosed using “Hanifin/Rajka criteria.”Hanifin/Rajka diagnostic criteria are described in Acta Derm Venereol(Stockh) 1980; Suppl 92:44-7. To establish a diagnosis of atopicdermatitis the patient requires the presence of at least 3 “basicfeatures” and 3 or more minor features listed below. The basic featuresinclude pruritus, typical morphology and distribution such as flexurallichenification or linearity, chronic or chronically-relapsingdermatitis, and personal or family history of atopy, such as asthma,allergic rhinitis, atopic dermatitis. The minor features includexerosis, ichthyosis, palmar hyperlinearity, or keratosis pilaris,immediate (type 1) skin-test reactivity, elevated serum IgE, early ageof onset, tendency toward cutaneous infections (especially Staph. aureusand Herpes simplex), impaired cell-mediated immunity, tendency towardnon-specific hand or foot dermatitis, nipple eczema, cheilitis,recurrent conjunctivitis, Dennie-Morgan infraorbital fold, keratoconus,anterior subcapsular cataracts, orbital darkening, facial pallor/facialerythema, pityriasis alba, anterior neck folds, and itch when sweating.Additional minor criteria include intolerance to wool and lipidsolvents, perifollicular accentuation, food intolerance, courseinfluenced by environmental or emotional factors, and whitedermographism/delayed blanch.

The severity of atopic dermatitis can be determined by “Rajka/Langelandcriteria,” as described in Rajka G and Langeland T, Acta Derm Venereol(Stockh) 1989; 144(Suppl):13-4. Three disease severity assessmentcategories are scored 1 to 3: I) extent of the body area involved, II)course, e.g., more or less than 3 months during one year or continuouscourse, and III) intensity, ranging from mild itch to severe itch,usually disturbing night's sleep. Scores of 1.5 or 2.5 are allowed.Overall disease severity is determined by the sum of individual scoresfrom the three disease assessment categories and the severity isdetermined by the sum of these scores with mild defined as a total scoreof 3-4, moderate as score of 4.5-7.5, and severe as a total score of8-9.

The term “Atopic Dermatitis Disease Severity Outcome Measure” or“ADDSOM” means a determination of certain signs, symptoms, features orparameters that have been associated with atopic dermatitis and that canbe quantitatively or qualitatively assessed. Exemplary ADDSOMs include,but are not limited to, “Eczema Area and Severity Index” (EAST),“Severity Scoring of Atopic Dermatitis” (SCORAD), “Investigator GlobalAssessment” (IGA), “Investigator Global Assessment of Signs” (IGSA),Rajka/Langeland Atopic Dermatitis Severity Score, and Patient-ReportedOutcomes including, but not limited to, Pruritus Visual Analog Scale (anaspect of disease severity assessed as part of SCORAD), Sleep LossVisual Analog Scale (an aspect of disease severity assessed as part ofSCORAD), Atopic Dermatitis Symptom Diary (ADSD), Atopic DermatitisImpact Questionnaire (ADIQ), Dermatology Life Quality Index (DLQI)(Finlay and Khan, Clin Exper Dermatol 1994;19:210), and 5-D Itch Scale(Elman et al., Br J Dermatol 2010;162(3):587-593).

The “Eczema Area and Severity Index” or “EAST” is a validated measureused in clinical settings to assess the severity and extent of AD,Hanifin et al., Exp Dermatol 2001; 10:11-18. Four individual bodyregions are assessed by a clinician or other medical professional: headand neck (H&N), upper limbs (UL; includes the external axillae andhands), trunk (includes the internal axillae and groin), and lower limbs(LL; includes the buttocks and feet). For each body region, the affectedbody surface area (BSA) is assessed and assigned a score of 0 to 6 (oroptionally 0-7 where 0 equals no eruption) for the percentage ofaffected BSA (0%-100%); each region is individually rated for theaverage degree of severity (0-3: none, mild, moderate, severe), withhalf steps allowed, for each of four clinical signs: erythema,induration-papulation, excoriations, and lichenification. A summed scoreof 0 to 12 is assigned to each body region; a total body region score isassigned based on the sum of the individual clinical signs score(maximum=12)×affected area score (maximum=6)×the body-region index(H&N-0.1, UL-0.2, trunk-0.3, LL-0.4). A total score (0-72) is assignedbased on the sum of total scores for each of the four body regionscores.

The “Investigator Global Assessment” or “IGA” is an assessment measureused in clinical settings to determine the severity of AD and clinicalresponse to treatment based on a five-point scale. A score of 0 (clear)means there are no inflammatory signs of atopic dermatitis and a scoreof 1 (almost clear) means there is just perceptible erythema and justperceptible papulation induration. Scores of 2, 3, or 4 (mild, moderate,severe) are based on the severity of erythema, papulation induration,oozing and crusting. The “Investigator Global Assessment of Signs” or“IGSA” uses a lesional assessment grade ranging from clear to severebased on an evaluation of erythema, edema, lichenified plaques orpapules, and excoriations. In addition, the assessed lesional grade maybe upgraded or downgraded based on the extent and location of the skinlesion.

The “Severity Scoring of Atopic Dermatitis” or “SCORAD” is a clinicalassessment of the severity of AD developed by the European Task Force onAtopic Dermatitis (consensus report, Dermatology 1993; 186:23-31). Threeaspects of disease severity are scored: (i) the extent of body areaaffected by inflammation with score assessed between 0-100, assigned as“A” in the overall total score, (ii) the intensity of six clinicalsigns, erythema, edema/population, oozing/crusting, excoriation,lichenification, and dryness, each assigned a score of 0-3 based onseverity (absent, mild, moderate, severe) for a total score ranging from0-18, assigned as “B” in the overall total score, and (iii) twosubjective measures that use patient-reported outcomes, the pruritusvisual analog score (ranging from 0 [no itch] to 10 [the worstimaginable itch]) and the sleep loss visual analog score (ranging from 0[no sleep loss] to 10 [the worst imaginable sleep loss]), each theaverage of the last three days or nights, assigned as “C” in the overalltotal score. The overall total score (0-103) is determined according tothe formula: A/5+(7B/2)+C.

The term “Patient-Reported Outcome” or “PRO” means a validatedquestionnaire or tool used in clinical practice or in clinical trials toassess quality of life from a patient's point of view regarding thepatient's atopic dermatitis disease symptoms which may include emotionaland functional impacts of the disease. A PRO is a report of the statusof a patient's health condition that comes directly from the patient,without interpretation of the patient's response by a clinician oranyone else. The outcome can be measured in absolute terms (e.g.,severity of a symptom, sign, or state of a disease) or as a change froma previous measure. In clinical trials, a PRO instrument can be used tomeasure the effect of a medical intervention on one or more concepts(i.e., the thing being measured, such as a symptom or group of symptoms,effects on a particular function or group of functions, or a group ofsymptoms or functions shown to measure the severity of a healthcondition). Exemplary PRO tools used to assess AD include, but are notlimited to, Pruritus Visual Analog Scale (an aspect of disease severityassessed as part of SCORAD), Sleep Loss Visual Analog Scale (an aspectof disease severity assessed as part of SCORAD), Atopic DermatitisSymptom Diary (ADSD), Atopic Dermatitis Impact Questionnaire (ADIQ),Dermatology Life Quality Index (DLQI) (Finlay and Khan, Clin ExperDermatol 1994;19:210), and 5-D Itch Scale (Elman et al., Br J Dermatol2010;162(3):587-593).

Compositions and Methods

The invention is based, at least in part, on the surprising andunexpected discovery that an anti-IL-13 antagonist monoclonal antibody,lebrikizumab, provides therapeutic benefit when administered to atopicdermatitis patients using the dosing regimens provided herein, includingpatients using topical corticosteroids concomitantly, as assessed byseveral efficacy outcome measures. Accordingly, provided herein aremethods of treating atopic dermatitis with IL-13 antagonists, includinganti-IL-13 antibodies such as lebrikizumab.

Lebrikizumab is a humanized, monoclonal, IgG4 antibody that binds IL-13with high affinity and blocks signaling through the activeIL-4Ralpha/IL-13Ralpha1 heterodimer. Lebrikizumab has been clinicallytested in asthma at various doses and the results have been reported inthe literature as summarized below.

In the MILLY study, adults with poorly controlled asthma despite inhaledcorticosteroid treatment were administered 250 mg of lebrikizumab orplacebo once every four weeks for a total of six months. Corren et al.,N Engl J Med 2011;365:1088-98. Lebrikizumab-treated patients,particularly those in the biomarker high (serum periostin-high)subgroup, exhibited greater improvements in lung function, as measuredby FEV₁, than those in the placebo group. Corren et al., supra.

The MOLLY study was a dose-ranging study in which asthma patients notreceiving inhaled corticosteroids received 125 mg of lebrikizumab or 250mg lebrikizumab or 500 mg lebrikizumab or placebo once every four weeksfor a total of 12 weeks. Noonan et al., J Allergy Clin Immunol2013;132:567-74. Although the mean relative change in FEV₁ wasnumerically higher in all lebrikizumab dose groups compared to theplacebo group, these differences were neither statistically norclinically significant. Noonan et al., supra. In addition, the MOLLYstudy did not meet the objectives for demonstrating a dose-response.Noonan et al., supra. One conclusion from the results is that blocking asingle cytokine, IL-13, in that population of asthma patients in theMOLLY study, was insufficient to improve lung function as measured byFEV₁ compared with existing treatments. Noonan et al., supra.

In the LUTE and VERSE studies, three different doses of lebrikizumab,37.5 mg, 125 mg and 250 mg, administered once every four weeks, weretested in moderate-to-severe asthmatic patients not controlled oninhaled corticosteroids. Hanania et al., Thorax 2015;70:748-756.Treatment with lebrikizumab reduced the rate of asthma exacerbations,which was more pronounced in the biomarker-high (serum periostin-high)patients (all doses: 60% reduction) than in the biomarker-low (serumperiostin-low) patients (all doses: 5% reduction); no cleardose—response was evident, however. Hanania et al., 2015, supra. Lungfunction also improved following lebrikizumab treatment, with greatestincrease in FEV₁ in periostin-high patients compared with periostin-lowpatients. Hanania et al., 2015, supra.

As described herein, both AD and asthma patients have elevated levels ofbiomarkers associated with IL-13 biology and Th-2 mediatedhypersensitivity. Accordingly, it was thought that the dose levels oflebrikizumab producing clinical benefit in asthma could also producebiological activity in AD patients. Thus, the initial doses proposed fortesting in AD patients as set forth in Clinical Study I and ClinicalStudy II described in the Examples were based on the published clinicalexperience with asthma as described above.

After initiation of Clinical Study I and Clinical Study II describedherein, the results of the phase 3 clinical studies of lebrikizumab inasthma (LAVOLTA I and LAVOLTA II) were analyzed and published onlineSep. 5, 2016 in Hanania et al., Lancet Respir Med 2016, available atdx(dot)doi(dot)org(slash)52213-2600(16)30265-X . LAVOLTA I and II werereplicate phase 3 studies to assess the efficacy and safety oflebrikizumab in patients with uncontrolled asthma despite inhaledcorticosteroids and at least one second controller. These phase 3studies were designed for the purpose of seeking marketing approval fromhealth authorities world-wide provided there was a demonstration ofstatistically significant efficacy in both studies and an acceptablesafety profile.

In LAVOLTA I and II, patients were treated with 37.5 mg lebrikizumab or125 mg lebrikizumab or placebo, administered once every four weeks, over52 weeks. The primary efficacy endpoint was a reduction in the rate ofexacerbations. Patients were stratified according to biomarker-high(serum periostin-high or blood eosinophil count-high) or biomarker-low(serum periostin-low or blood eosinophil count-low). As reported inHanania et al., 2016, supra, the efficacy results across both studieswere inconsistent. Lebrikizumab significantly reduced the rate ofexacerbations in biomarker-high patients in LAVOLTA I but not in LAVOLTAII. Hanania et al., 2016, supra. Both studies failed to show a cleardose-response although pharmacokinetic and pharmacodynamics results wereconsistent with those from the phase II studies described above,indicating that drug exposures were similar and that the IL-13 pathwaywas inhibited. Hanania et al., 2016, supra.

Accordingly, although the lebrikizumab dosing regimens tested in theatopic dermatitis Clinical Study I and Clinical Study II describedherein were based on the clinical experience with asthma, it wasuncertain whether lebrikizumab would provide clinically meaningfulbenefit to atopic dermatitis patients at the dosing regimens describedfor Clinical Study I and Clinical Study II. The inconsistent results oflebrikizumab in asthma patients and the lack of a clear dose-responseacross multiple studies as summarized above, including in view of theresults reported for the phase 3 LAVOLTA I and II studies, contribute tosuch uncertainty with respect to therapeutic benefit of lebrikizumab inatopic dermatitis patients. Prior to the invention described herein, itwould have been unpredictable whether lebrikizumab could provideclinically meaningful benefit to atopic dermatitis patients at thedosing regimens provided herein.

Exemplary Antibodies Anti-IL-13 Antibodies

In one aspect, the invention provides isolated antibodies that bind tohuman IL-13.

Exemplary anti-IL-13 antibodies are known and include, for example, butnot limited to, lebrikizumab, IMA-026, IMA-638 (also referred to as,anrukinzumab, INN No. 910649-32-0; QAX-576), tralokinumab (also referredto as CAT-354, CAS No. 1044515-88-9); AER-001, ABT-308 (also referred toas humanized 13C5.5 antibody. Examples of such anti-IL-13 antibodies andother inhibitors of IL-13 are disclosed, for example, in WO 2005/062967,WO2008/086395, WO2006/085938, U.S. Pat. No. 7,615,213, U.S. Pat. No.7,501,121, WO2007/036745, WO2010/073119, WO2007/045477, WO 2014/165771.In one embodiment, the anti-IL-13 antibody is a humanized IgG4 antibody.In one embodiment, the anti-IL-13 antibody is lebrikizumab. In oneembodiment, the anti-IL-13 antibody comprises three heavy chain HVRs,HVR-H1 (SEQ ID NO.: 5), HVR-H2 (SEQ ID NO.: 6), and HVR-H3 (SEQ ID NO.:7). In one embodiment, the anti-IL-13 antibody comprises three lightchain HVRS, HVR-L1 (SEQ ID NO.: 8), HVR-L2 (SEQ ID NO.: 9), and HVR-L3(SEQ ID NO.: 10). In one embodiment, the anti-IL-13 antibody comprisesthree heavy chain HVRs and three light chain HVRs, HVR-H1 (SEQ ID NO.:5), HVR-H2 (SEQ ID NO.: 6), HVR-H3 (SEQ ID NO.: 7), HVR-L1 (SEQ ID NO.:8), HVR-L2 (SEQ ID NO.: 9), and HVR-L3 (SEQ ID NO.: 10). In oneembodiment, the anti-IL-13 antibody comprises a variable heavy chainregion, VH, having an amino acid sequence selected from SEQ ID NOs. 1,3, and 24. In one embodiment, the anti-IL-13 antibody comprises avariable light chain region, VL, having an amino acid sequence selectedfrom SEQ ID NOs.: 2, 4, and 25. In one embodiment, the anti-IL-13antibody comprises a variable heavy chain region, VH, having an aminoacid sequence selected from SEQ ID NOs. 1, 3, and 24 and a variablelight chain region, VL, having an amino acid sequence selected from SEQID NOs.: 2, 4, and 25.

In another embodiment, the antibody comprises the variable regionsequences SEQ ID NO:1 and SEQ ID NO:2. In another embodiment, theantibody comprises the variable region sequences SEQ ID NO:1 and SEQ IDNO:4. In another embodiment, the antibody comprises the variable regionsequences SEQ ID NO:1 and SEQ ID NO:25. In another embodiment, theantibody comprises the variable region sequences SEQ ID NO:3 and SEQ IDNO:2. In another embodiment, the antibody comprises the variable regionsequences SEQ ID NO:3 and SEQ ID NO:4. In another embodiment, theantibody comprises the variable region sequences SEQ ID NO:3 and SEQ IDNO:25. In another embodiment, the antibody comprises the variable regionsequences SEQ ID NO:24 and SEQ ID NO:2. In another embodiment, theantibody comprises the variable region sequences SEQ ID NO:24 and SEQ IDNO:4. In another embodiment, the antibody comprises the variable regionsequences SEQ ID NO:24 and SEQ ID NO:25.

In any of the above embodiments, an anti-IL-13 antibody can behumanized. In one embodiment, an anti-IL-13 antibody comprises HVRs asin any of the above embodiments, and further comprises an acceptor humanframework, e.g. a human immunoglobulin framework or a human consensusframework.

In another aspect, an anti-IL-13 antibody comprises a heavy chainvariable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acidsequence of SEQ ID NO:1. In certain embodiments, a VH sequence having atleast 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identitycontains substitutions (e.g., conservative substitutions), insertions,or deletions relative to the reference sequence, but an anti-IL-13antibody comprising that sequence retains the ability to bind to humanIL-13. In certain embodiments, a total of 1 to 10 amino acids have beensubstituted, altered inserted and/or deleted in SEQ ID NO: 1. In certainembodiments, substitutions, insertions, or deletions occur in regionsoutside the HVRs (i.e., in the FRs). Optionally, the anti-IL-13 antibodycomprises the VH sequence in SEQ ID NO: 1, including post-translationalmodifications of that sequence. Optionally, the anti-IL-13 antibodycomprises the VH sequence in SEQ ID NO: 3, including post-translationalmodifications of that sequence. Optionally, the anti-IL-13 antibodycomprises the VH sequence in SEQ ID NO: 24, including post-translationalmodifications of that sequence.

In another aspect, an anti-IL-13 antibody is provided, wherein theantibody comprises a light chain variable domain (VL) having at least90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequenceidentity to the amino acid sequence of SEQ ID NO:2. In certainembodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, or 99% identity contains substitutions (e.g.,conservative substitutions), insertions, or deletions relative to thereference sequence, but an anti-IL-13 antibody comprising that sequenceretains the ability to bind to IL-13. In certain embodiments, a total of1 to 10 amino acids have been substituted, inserted and/or deleted inSEQ ID NO:2. In certain embodiments, the substitutions, insertions, ordeletions occur in regions outside the HVRs (i.e., in the FRs).Optionally, the anti-IL-13 antibody comprises the VL sequence in SEQ IDNO:2, including post-translational modifications of that sequence.

Optionally, the anti-IL-13 antibody comprises the VL sequence in SEQ IDNO: 4, including post-translational modifications of that sequence.Optionally, the anti-IL-13 antibody comprises the VL sequence in SEQ IDNO: 25, including post-translational modifications of that sequence.

In another aspect, an anti-IL-13 antibody is provided, wherein theantibody comprises a VH as in any of the embodiments provided above, anda VL as in any of the embodiments provided above.

In a further aspect, the invention provides an antibody that binds tothe same epitope as an anti-IL-13 antibody provided herein. For example,in certain embodiments, an antibody is provided that binds to the sameepitope as or can by competitively inhibited by an anti-IL-13 antibodycomprising a VH sequence of SEQ ID NO:1 and a VL sequence of SEQ IDNO:2.

In a further aspect of the invention, an anti-IL-13 antibody accordingto any of the above embodiment can be a monoclonal antibody, including achimeric, humanized or human antibody. In one embodiment, an anti-IL-13antibody is an antibody fragment, e.g., a Fv, Fab, Fab′, scFv, diabody,or F(ab′)2 fragment. In another embodiment, the antibody is a fulllength antibody, e.g., an intact IgG1 or IgG4 antibody or other antibodyclass or isotype as defined herein. According to another embodiment, theantibody is a bispecific antibody. In one embodiment, the bispecificantibody comprises the HVRs or comprises the VH and VL regions describedabove.

In one embodiment, the anti-IL-13 antibody comprises three heavy chainHVRs, HVR-H1 (SEQ ID NO.: 13), HVR-H2 (SEQ ID NO.: 14), and HVR-H3 (SEQID NO.: 15). In one embodiment, the anti-IL-13 antibody comprises threelight chain HVRS, HVR-L1 (SEQ ID NO.: 16), HVR-L2 (SEQ ID NO.: 17), andHVR-L3 (SEQ ID NO.: 18). In one embodiment, the anti-IL-13 antibodycomprises three heavy chain HVRs and three light chain HVRs, HVR-H1 (SEQID NO.: 13), HVR-H2 (SEQ ID NO.: 14), HVR-H3 (SEQ ID NO.: 15), HVR-L1(SEQ ID NO.: 16), HVR-L2 (SEQ ID NO.: 17), and HVR-L3 (SEQ ID NO.: 18).In one embodiment, the anti-IL-13 antibody comprises a variable heavychain region, VH, having the amino acid sequence of SEQ ID NO: 12. Inone embodiment, the anti-IL-13 antibody comprises a variable light chainregion, VL, having the amino acid sequence of SEQ ID NO: 11. In oneembodiment, the anti-IL-13 antibody comprises a variable heavy chainregion, VH, having the amino acid sequence of SEQ ID NO: 12 and avariable light chain region, VL, having the amino acid sequence of SEQID NO: 11.

In any of the above embodiments, an anti-IL-13 antibody can behumanized. In one embodiment, an anti-IL-13 antibody comprises HVRs asin any of the above embodiments, and further comprises an acceptor humanframework, e.g. a human immunoglobulin framework or a human consensusframework.

In another aspect, an anti-IL-13 antibody comprises a heavy chainvariable domain (VH) sequence having at least 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acidsequence of SEQ ID NO:12. In certain embodiments, a VH sequence havingat least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identitycontains substitutions (e.g., conservative substitutions), insertions,or deletions relative to the reference sequence, but an anti-IL-13antibody comprising that sequence retains the ability to bind to humanIL-13. In certain embodiments, a total of 1 to 10 amino acids have beensubstituted, altered inserted and/or deleted in SEQ ID NO: 12. Incertain embodiments, substitutions, insertions, or deletions occur inregions outside the HVRs (i.e., in the FRs). Optionally, the anti-IL-13antibody comprises the VH sequence in SEQ ID NO: 12, includingpost-translational modifications of that sequence.

In another aspect, an anti-IL-13 antibody is provided, wherein theantibody comprises a light chain variable domain (VL) having at least90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequenceidentity to the amino acid sequence of SEQ ID NO:11. In certainembodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, or 99% identity contains substitutions (e.g.,conservative substitutions), insertions, or deletions relative to thereference sequence, but an anti-IL-13 antibody comprising that sequenceretains the ability to bind to IL-13. In certain embodiments, a total of1 to 10 amino acids have been substituted, inserted and/or deleted inSEQ ID NO:11. In certain embodiments, the substitutions, insertions, ordeletions occur in regions outside the HVRs (i.e., in the FRs).Optionally, the anti-IL-13 antibody comprises the VL sequence in SEQ IDNO:11, including post-translational modifications of that sequence.

In another aspect, an anti-IL-13 antibody is provided, wherein theantibody comprises a VH as in any of the embodiments provided above, anda VL as in any of the embodiments provided above.

In a further aspect, the invention provides an antibody that binds tothe same epitope as an anti-IL-13 antibody provided herein. For example,in certain embodiments, an antibody is provided that binds to the sameepitope as or can by competitively inhibited by an anti-IL-13 antibodycomprising a VH sequence of SEQ ID NO:12 and a VL sequence of SEQ IDNO:11.

In a further aspect of the invention, an anti-IL-13 antibody accordingto any of the above embodiment can be a monoclonal antibody, including achimeric, humanized or human antibody. In one embodiment, an anti-IL-13antibody is an antibody fragment, e.g., a Fv, Fab, Fab′, scFv, diabody,or F(ab′)2 fragment. In another embodiment, the antibody is a fulllength antibody, e.g., an intact IgG1 or IgG4 antibody or other antibodyclass or isotype as defined herein. According to another embodiment, theantibody is a bispecific antibody. In one embodiment, the bispecificantibody comprises the HVRs or comprises the VH and VL regions describedabove.

In a further aspect, an anti-IL-13 antibody according to any of theabove embodiments may incorporate any of the features, singly or incombination, as described in Sections 1-7 below:

1. Antibody Affinity

In certain embodiments, an antibody provided herein has a dissociationconstant (Kd) of ≤1 μM, ≤100 nM, ≤10 nM, ≤1 nM, ≤0.1 nM, ≤0.01 nM, or≤0.001 nM (e.g. 10-8 M or less, e.g. from 10-8 M to 10-13 M, e.g., from10-9 M to 10-13 M).

In one embodiment, Kd is measured by a radiolabeled antigen bindingassay (MA) performed with the Fab version of an antibody of interest andits antigen as described by the following assay. Solution bindingaffinity of Fabs for antigen is measured by equilibrating Fab with aminimal concentration of (125I)-labeled antigen in the presence of atitration series of unlabeled antigen, then capturing bound antigen withan anti-Fab antibody-coated plate (see, e.g., Chen et al., J. Mol. Biol.293:865-881(1999)). To establish conditions for the assay, MICROTITER®multi-well plates (Thermo Scientific) are coated overnight with 5 μg/mlof a capturing anti-Fab antibody (Cappel Labs) in 50 mM sodium carbonate(pH 9.6), and subsequently blocked with 2% (w/v) bovine serum albumin inPBS for two to five hours at room temperature (approximately 23° C.). Ina non-adsorbent plate (Nunc #269620), 100 μM or 26 μM [125I]-antigen aremixed with serial dilutions of a Fab of interest (e.g., consistent withassessment of the anti-VEGF antibody, Fab-12, in Presta et al., CancerRes. 57:4593-4599 (1997)). The Fab of interest is then incubatedovernight; however, the incubation may continue for a longer period(e.g., about 65 hours) to ensure that equilibrium is reached.Thereafter, the mixtures are transferred to the capture plate forincubation at room temperature (e.g., for one hour). The solution isthen removed and the plate washed eight times with 0.1% polysorbate 20(TWEEN-20®) in PBS. When the plates have dried, 150 μl/well ofscintillant (MICROSCINT-20 TM; Packard) is added, and the plates arecounted on a TOPCOUNT TM gamma counter (Packard) for ten minutes.Concentrations of each Fab that give less than or equal to 20% ofmaximal binding are chosen for use in competitive binding assays.

According to another embodiment, Kd is measured using surface plasmonresonance assays using a BIACORE®-2000 or a BIACORE ®-3000 (BlAcore,Inc., Piscataway, N.J.) at 25° C. with immobilized antigen CMS chips at˜10 response units (RU). Briefly, carboxymethylated dextran biosensorchips (CMS, BIACORE, Inc.) are activated withN-ethyl-N′-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) andN-hydroxysuccinimide (NETS) according to the supplier's instructions.Antigen is diluted with 10 mM sodium acetate, pH 4.8, to 5 μg/ml (−0.2μM) before injection at a flow rate of 5 μl/minute to achieveapproximately 10 response units (RU) of coupled protein. Following theinjection of antigen, 1 M ethanolamine is injected to block unreactedgroups. For kinetics measurements, two-fold serial dilutions of Fab(0.78 nM to 500 nM) are injected in PBS with 0.05% polysorbate 20(TWEEN-20TM) surfactant (PBST) at 25° C. at a flow rate of approximately25 μl/min. Association rates (kon) and dissociation rates (koff) arecalculated using a simple one-to-one Langmuir binding model (BIACORE ®Evaluation Software version 3.2) by simultaneously fitting theassociation and dissociation sensorgrams. The equilibrium dissociationconstant (Kd) is calculated as the ratio koff/kon. See, e.g., Chen etal., J. Mol. Biol. 293:865-881 (1999). If the on-rate exceeds 106 M-1s-1 by the surface plasmon resonance assay above, then the on-rate canbe determined by using a fluorescent quenching technique that measuresthe increase or decrease in fluorescence emission intensity(excitation=295 nm; emission=340 nm, 16 nm band-pass) at 25° C. of a 20nM antigen antibody (Fab form) in PBS, pH 7.2, in the presence ofincreasing concentrations of antigen as measured in a spectrometer, suchas a stop-flow equipped spectrophotometer (Aviv Instruments) or a8000-series SLM-AMINCO TM spectrophotometer (ThermoSpectronic) with astirred cuvette.

2. Antibody Fragments

In certain embodiments, an antibody provided herein is an antibodyfragment. Antibody fragments include, but are not limited to, Fab, Fab′,Fab′-SH, F(ab′)2, Fv, and scFv fragments, and other fragments describedbelow. For a review of certain antibody fragments, see Hudson et al.Nat. Med. 9:129-134 (2003). For a review of scFv fragments, see, e.g.,Pluckthiin, in The Pharmacology of Monoclonal Antibodies, vol. 113,Rosenburg and Moore eds., (Springer-Verlag, New York), pp. 269-315(1994); see also WO 93/16185; and U.S. Pat. Nos. 5,571,894 and5,587,458. For discussion of Fab and F(ab′)2 fragments comprisingsalvage receptor binding epitope residues and having increased in vivohalf-life, see U.S. Pat. No. 5,869,046.

Diabodies are antibody fragments with two antigen-binding sites that maybe bivalent or bispecific. See, for example, EP 404,097; WO 1993/01161;Hudson et al., Nat. Med. 9:129-134 (2003); and Hollinger et al., Proc.Natl. Acad. Sci. USA 90: 6444-6448 (1993). Triabodies and tetrabodiesare also described in Hudson et al., Nat. Med. 9:129-134 (2003).

Single-domain antibodies are antibody fragments comprising all or aportion of the heavy chain variable domain or all or a portion of thelight chain variable domain of an antibody. In certain embodiments, asingle-domain antibody is a human single-domain antibody (Domantis,Inc., Waltham, Mass.; see, e.g., U.S. Pat. No. 6,248,516 B1).

Antibody fragments can be made by various techniques, including but notlimited to proteolytic digestion of an intact antibody as well asproduction by recombinant host cells (e.g. E. coli or phage), asdescribed herein.

3. Chimeric and Humanized Antibodies

In certain embodiments, an antibody provided herein is a chimericantibody. Certain chimeric antibodies are described, e.g., in U.S. Pat.No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA,81:6851-6855 (1984)). In one example, a chimeric antibody comprises anon-human variable region (e.g., a variable region derived from a mouse,rat, hamster, rabbit, or non-human primate, such as a monkey) and ahuman constant region. In a further example, a chimeric antibody is a“class switched” antibody in which the class or subclass has beenchanged from that of the parent antibody. Chimeric antibodies includeantigen-binding fragments thereof.

In certain embodiments, a chimeric antibody is a humanized antibody.Typically, a non-human antibody is humanized to reduce immunogenicity tohumans, while retaining the specificity and affinity of the parentalnon-human antibody. Generally, a humanized antibody comprises one ormore variable domains in which HVRs, e.g., CDRs, (or portions thereof)are derived from a non-human antibody, and FRs (or portions thereof) arederived from human antibody sequences. A humanized antibody optionallywill also comprise at least a portion of a human constant region. Insome embodiments, some FR residues in a humanized antibody aresubstituted with corresponding residues from a non-human antibody (e.g.,the antibody from which the HVR residues are derived), e.g., to restoreor improve antibody specificity or affinity.

Humanized antibodies and methods of making them are reviewed, e.g., inAlmagro and Fransson, Front. Biosci. 13:1619-1633 (2008), and arefurther described, e.g., in Riechmann et al., Nature 332:323-329 (1988);Queen et al., Proc. Nat'l Acad. Sci. USA 86:10029-10033 (1989); U.S.Pat. Nos. 5, 821,337, 7,527,791, 6,982,321, and 7,087,409; Kashmiri etal., Methods 36:25-34 (2005) (describing SDR (a-CDR) grafting); Padlan,Mol. Immunol. 28:489-498 (1991) (describing “resurfacing”); Dall'Acquaet al., Methods 36:43-60 (2005) (describing “FR shuffling”); and Osbournet al., Methods 36:61-68 (2005) and Klimka et al., Br. J. Cancer,83:252-260 (2000) (describing the “guided selection” approach to FRshuffling).

Human framework regions that may be used for humanization include butare not limited to: framework regions selected using the “best-fit”method (see, e.g., Sims et al. J. Immunol. 151:2296 (1993)); frameworkregions derived from the consensus sequence of human antibodies of aparticular subgroup of light or heavy chain variable regions (see, e.g.,Carter et al. Proc. Natl. Acad. Sci. USA, 89:4285 (1992); and Presta etal. J. Immunol., 151:2623 (1993)); human mature (somatically mutated)framework regions or human germline framework regions (see, e.g.,Almagro and Fransson, Front. Biosci. 13:1619-1633 (2008)); and frameworkregions derived from screening FR libraries (see, e.g., Baca et al., J.Biol. Chem. 272:10678-10684 (1997) and Rosok et al., J. Biol. Chem.271:22611-22618 (1996)).

4. Human Antibodies

In certain embodiments, an antibody provided herein is a human antibody.Human antibodies can be produced using various techniques known in theart. Human antibodies are described generally in van Dijk and van deWinkel, Curr. Opin. Pharmacol. 5: 368-74 (2001) and Lonberg, Curr. Opin.Immunol. 20:450-459 (2008).

Human antibodies may be prepared by administering an immunogen to atransgenic animal that has been modified to produce intact humanantibodies or intact antibodies with human variable regions in responseto antigenic challenge. Such animals typically contain all or a portionof the human immunoglobulin loci, which replace the endogenousimmunoglobulin loci, or which are present extrachromosomally orintegrated randomly into the animal's chromosomes. In such transgenicmice, the endogenous immunoglobulin loci have generally beeninactivated. For review of methods for obtaining human antibodies fromtransgenic animals, see Lonberg, Nat. Biotech. 23:1117-1125 (2005). Seealso, e.g., U.S. Pat. Nos. 6,075,181 and 6,150,584 describingXENOMOUSETM technology; U.S. Pat. No. 5,770,429 describing HUMAB®technology; U.S. Pat. No. 7,041,870 describing K-M MOUSE® technology,and U.S. Patent Application Publication No. US 2007/0061900, describingVELOCIMOUSE® technology). Human variable regions from intact antibodiesgenerated by such animals may be further modified, e.g., by combiningwith a different human constant region.

Human antibodies can also be made by hybridoma-based methods. Humanmyeloma and mouse-human heteromyeloma cell lines for the production ofhuman monoclonal antibodies have been described. (See, e.g., Kozbor J.Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal AntibodyProduction Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc.,New York, 1987); and Boerner et al., J. Immunol., 147: 86 (1991).) Humanantibodies generated via human B-cell hybridoma technology are alsodescribed in Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562(2006). Additional methods include those described, for example, in U.S.Pat. No. 7,189,826 (describing production of monoclonal human IgMantibodies from hybridoma cell lines) and Ni, Xiandai Mianyixue,26(4):265-268 (2006) (describing human-human hybridomas). Humanhybridoma technology (Trioma technology) is also described in Vollmersand Brandlein, Histology and Histopathology, 20(3):927-937 (2005) andVollmers and Brandlein, Methods and Findings in Experimental andClinical Pharmacology, 27(3):185-91 (2005).

Human antibodies may also be generated by isolating Fv clone variabledomain sequences selected from human-derived phage display libraries.Such variable domain sequences may then be combined with a desired humanconstant domain. Techniques for selecting human antibodies from antibodylibraries are described below.

5. Library-Derived Antibodies

Antibodies of the invention may be isolated by screening combinatoriallibraries for antibodies with the desired activity or activities. Forexample, a variety of methods are known in the art for generating phagedisplay libraries and screening such libraries for antibodies possessingthe desired binding characteristics. Such methods are reviewed, e.g., inHoogenboom et al. in Methods in Molecular Biology 178:1-37 (O'Brien etal., ed., Human Press, Totowa, N.J., 2001) and further described, e.g.,in the McCafferty et al., Nature 348:552-554; Clackson et al., Nature352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992);Marks and Bradbury, in Methods in Molecular Biology 248:161-175 (Lo,ed., Human Press, Totowa, N.J., 2003); Sidhu et al., J. Mol. Biol.338(2): 299-310 (2004); Lee et al., J. Mol. Biol. 340(5): 1073-1093(2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467-12472(2004); and Lee et al., J. Immunol. Methods 284(1-2): 119-132(2004).

In certain phage display methods, repertoires of VH and VL genes areseparately cloned by polymerase chain reaction (PCR) and recombinedrandomly in phage libraries, which can then be screened forantigen-binding phage as described in Winter et al., Ann. Rev. Immunol.,12: 433-455 (1994). Phage typically display antibody fragments, eitheras single-chain Fv (scFv) fragments or as Fab fragments. Libraries fromimmunized sources provide high-affinity antibodies to the immunogenwithout the requirement of constructing hybridomas. Alternatively, thenaive repertoire can be cloned (e.g., from human) to provide a singlesource of antibodies to a wide range of non-self and also self antigenswithout any immunization as described by Griffiths et al., EMBO J, 12:725-734 (1993). Finally, naive libraries can also be made syntheticallyby cloning unrearranged V-gene segments from stem cells, and using PCRprimers containing random sequence to encode the highly variable CDR3regions and to accomplish rearrangement in vitro, as described byHoogenboom and Winter, J. Mol. Biol., 227: 381-388 (1992). Patentpublications describing human antibody phage libraries include, forexample: U.S. Pat. No. 5,750,373, and U.S. Patent Publication Nos.2005/0079574, 2005/0119455, 2005/0266000, 2007/0117126, 2007/0160598,2007/0237764, 2007/0292936, and 2009/0002360.

Antibodies or antibody fragments isolated from human antibody librariesare considered human antibodies or human antibody fragments herein.

6. Multispecific Antibodies

In certain embodiments, an antibody provided herein is a multispecificantibody, e.g. a bispecific antibody. Multispecific antibodies aremonoclonal antibodies that have binding specificities for at least twodifferent sites. In certain embodiments, one of the bindingspecificities is for IL-13 and the other is for any other antigen. Incertain embodiments, bispecific antibodies may bind to two differentepitopes of IL-13. Bispecific antibodies may also be used to localizecytotoxic agents to cells. Bispecific antibodies can be prepared as fulllength antibodies or antibody fragments.

Techniques for making multispecific antibodies include, but are notlimited to, recombinant co-expression of two immunoglobulin heavychain-light chain pairs having different specificities (see Milstein andCuello, Nature 305: 537 (1983)), WO 93/08829, and Traunecker et al.,EMBO J. 10: 3655 (1991)), and “knob-in-hole” engineering (see, e.g.,U.S. Pat. No. 5,731,168). Multi-specific antibodies may also be made byengineering electrostatic steering effects for making antibodyFc-heterodimeric molecules (WO 2009/089004A1); cross-linking two or moreantibodies or fragments (see, e.g., U.S. Pat. No. 4,676,980, and Brennanet al., Science, 229: 81 (1985)); using leucine zippers to producebi-specific antibodies (see, e.g., Kostelny et al., J. Immunol.,148(5):1547-1553 (1992)); using “diabody” technology for makingbispecific antibody fragments (see, e.g., Hollinger et al., Proc. Natl.Acad. Sci. USA, 90:6444-6448 (1993)); and using single-chain Fv (sFv)dimers (see,e.g. Gruber et al., J. Immunol., 152:5368 (1994)); andpreparing trispecific antibodies as described, e.g., in Tutt et al. J.Immunol. 147: 60 (1991).

Engineered antibodies with three or more functional antigen bindingsites, including “Octopus antibodies,” are also included herein (see,e.g. US 2006/0025576A1).

The antibody or fragment herein also includes a “Dual Acting FAb” or“DAF” comprising an antigen binding site that binds to IL-13 as well asanother, different antigen (see, US 2008/0069820, for example).

7. Antibody Variants

In certain embodiments, amino acid sequence variants of the antibodiesprovided herein are contemplated. For example, it may be desirable toimprove the binding affinity and/or other biological properties of theantibody. Amino acid sequence variants of an antibody may be prepared byintroducing appropriate modifications into the nucleotide sequenceencoding the antibody, or by peptide synthesis. Such modificationsinclude, for example, deletions from, and/or insertions into and/orsubstitutions of residues within the amino acid sequences of theantibody. Any combination of deletion, insertion, and substitution canbe made to arrive at the final construct, provided that the finalconstruct possesses the desired characteristics, e.g., antigen-binding.

Substitution, Insertion, and Deletion Variants

In certain embodiments, antibody variants having one or more amino acidsubstitutions are provided. Sites of interest for substitutionalmutagenesis include the HVRs and FRs. Conservative substitutions areshown in Table 1 under the heading of “conservative substitutions.” Moresubstantial changes are provided in Table 1 under the heading of“exemplary substitutions,” and as further described below in referenceto amino acid side chain classes. Amino acid substitutions may beintroduced into an antibody of interest and the products screened for adesired activity, e.g., retained/improved antigen binding, decreasedimmunogenicity, or improved ADCC or CDC.

TABLE 1 Original Exemplary Conservative Residue SubstitutionsSubstitutions Ala (A) Val; Leu; Ile Val Arg (R) Lys; Gln; Asn Lys Asn(N) Gln; His; Asp, Lys; Arg Gln Asp (D) Glu; Asn Glu Cys (C) Ser; AlaSer Gln (Q) Asn; Glu Asn Glu (E) Asp; Gln Asp Gly (G) Ala Ala His (H)Asn; Gln; Lys; Arg Arg Ile (I) Leu; Val; Met; Ala; Phe; Norleucine LeuLeu (L) Norleucine; Ile; Val; Met; Ala; Phe Ile Lys (K) Arg; Gln; AsnArg Met (M) Leu; Phe; Ile Leu Phe (F) Trp; Leu; Val; Ile; Ala; Tyr TyrPro (P) Ala Ala Ser (S) Thr Thr Thr (T) Val; Ser Ser Trp (W) Tyr; PheTyr Tyr (Y) Trp; Phe; Thr; Ser Phe Val (V) Ile; Leu; Met; Phe; Ala;Norleucine Leu

Amino acids may be grouped according to common side-chain properties:

-   -   (1) hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile;    -   (2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln;    -   (3) acidic: Asp, Glu;    -   (4) basic: His, Lys, Arg;    -   (5) residues that influence chain orientation: Gly, Pro;    -   (6) aromatic: Trp, Tyr, Phe.

Non-conservative substitutions will entail exchanging a member of one ofthese classes for another class.

One type of substitutional variant involves substituting one or morehypervariable region residues of a parent antibody (e.g. a humanized orhuman antibody). Generally, the resulting variant(s) selected forfurther study will have modifications (e.g., improvements) in certainbiological properties (e.g., increased affinity, reduced immunogenicity)relative to the parent antibody and/or will have substantially retainedcertain biological properties of the parent antibody. An exemplarysubstitutional variant is an affinity matured antibody, which may beconveniently generated, e.g., using phage display-based affinitymaturation techniques such as those described herein. Briefly, one ormore HVR residues are mutated and the variant antibodies displayed onphage and screened for a particular biological activity (e.g. bindingaffinity).

Alterations (e.g., substitutions) may be made in HVRs, e.g., to improveantibody affinity. Such alterations may be made in HVR “hotspots,” i.e.,residues encoded by codons that undergo mutation at high frequencyduring the somatic maturation process (see, e.g., Chowdhury, MethodsMol. Biol. 207:179-196 (2008)), and/or SDRs (a-CDRs), with the resultingvariant VH or VL being tested for binding affinity. Affinity maturationby constructing and reselecting from secondary libraries has beendescribed, e.g., in Hoogenboom et al. in Methods in Molecular Biology178:1-37 (O'Brien et al., ed., Human Press, Totowa, N.J., (2001).) Insome embodiments of affinity maturation, diversity is introduced intothe variable genes chosen for maturation by any of a variety of methods(e.g., error-prone PCR, chain shuffling, or oligonucleotide-directedmutagenesis). A secondary library is then created. The library is thenscreened to identify any antibody variants with the desired affinity.Another method to introduce diversity involves HVR-directed approaches,in which several HVR residues (e.g., 4-6 residues at a time) arerandomized. HVR residues involved in antigen binding may be specificallyidentified, e.g., using alanine scanning mutagenesis or modeling. CDR-H3and CDR-L3 in particular are often targeted.

In certain embodiments, substitutions, insertions, or deletions mayoccur within one or more HVRs so long as such alterations do notsubstantially reduce the ability of the antibody to bind antigen. Forexample, conservative alterations (e.g., conservative substitutions asprovided herein) that do not substantially reduce binding affinity maybe made in HVRs. Such alterations may be outside of HVR “hotspots” orSDRs. In certain embodiments of the variant VH and VL sequences providedabove, each HVR either is unaltered, or contains no more than one, twoor three amino acid substitutions.

A useful method for identification of residues or regions of an antibodythat may be targeted for mutagenesis is called “alanine scanningmutagenesis” as described by Cunningham and Wells (1989) Science,244:1081-1085. In this method, a residue or group of target residues(e.g., charged residues such as arg, asp, his, lys, and glu) areidentified and replaced by a neutral or negatively charged amino acid(e.g., alanine or polyalanine) to determine whether the interaction ofthe antibody with antigen is affected. Further substitutions may beintroduced at the amino acid locations demonstrating functionalsensitivity to the initial substitutions. Alternatively, oradditionally, a crystal structure of an antigen-antibody complex toidentify contact points between the antibody and antigen. Such contactresidues and neighboring residues may be targeted or eliminated ascandidates for substitution. Variants may be screened to determinewhether they contain the desired properties.

Amino acid sequence insertions include amino- and/or carboxyl-terminalfusions ranging in length from one residue to polypeptides containing ahundred or more residues, as well as intrasequence insertions of singleor multiple amino acid residues. Examples of terminal insertions includean antibody with an N-terminal methionyl residue. Other insertionalvariants of the antibody molecule include the fusion to the N- orC-terminus of the antibody to an enzyme (e.g. for ADEPT) or apolypeptide which increases the serum half-life of the antibody.

Glycosylation Variants

In certain embodiments, an antibody provided herein is altered toincrease or decrease the extent to which the antibody is glycosylated.Addition or deletion of glycosylation sites to an antibody may beconveniently accomplished by altering the amino acid sequence such thatone or more glycosylation sites is created or removed.

Where the antibody comprises an Fc region, the carbohydrate attachedthereto may be altered. Native antibodies produced by mammalian cellstypically comprise a branched, biantennary oligosaccharide that isgenerally attached by an N-linkage to Asn297 of the CH2 domain of the Fcregion. See, e.g., Wright et al. TIBTECH 15:26-32 (1997). Theoligosaccharide may include various carbohydrates, e.g., mannose,N-acetyl glucosamine (GlcNAc), galactose, and sialic acid, as well as afucose attached to a GlcNAc in the “stem” of the biantennaryoligosaccharide structure. In some embodiments, modifications of theoligosaccharide in an antibody of the invention may be made in order tocreate antibody variants with certain improved properties.

In one embodiment, antibody variants are provided having a carbohydratestructure that lacks fucose attached (directly or indirectly) to an Fcregion. For example, the amount of fucose in such antibody may be from1% to 80%, from 1% to 65%, from 5% to 65% or from 20% to 40%. The amountof fucose is determined by calculating the average amount of fucosewithin the sugar chain at Asn297, relative to the sum of allglycostructures attached to Asn 297 (e. g. complex, hybrid and highmannose structures) as measured by MALDI-TOF mass spectrometry, asdescribed in WO 2008/077546, for example. Asn297 refers to theasparagine residue located at about position 297 in the Fc region (Eunumbering of Fc region residues); however, Asn297 may also be locatedabout ±3 amino acids upstream or downstream of position 297, i.e.,between positions 294 and 300, due to minor sequence variations inantibodies. Such fucosylation variants may have improved ADCC function.See, e.g., US Patent Publication Nos. US 2003/0157108 (Presta, L.); US2004/0093621 (Kyowa Hakko Kogyo Co., Ltd). Examples of publicationsrelated to “defucosylated” or “fucose-deficient” antibody variantsinclude: US 2003/0157108; WO 2000/61739; WO 2001/29246; US 2003/0115614;US 2002/0164328; US 2004/0093621; US 2004/0132140; US 2004/0110704; US2004/0110282; US 2004/0109865; WO 2003/085119; WO 2003/084570; WO2005/035586; WO 2005/035778; WO2005/053742; WO2002/031140; Okazaki etal. J. Mol. Biol. 336:1239-1249 (2004); Yamane-Ohnuki et al. Biotech.Bioeng. 87: 614 (2004). Examples of cell lines capable of producingdefucosylated antibodies include Lec13 CHO cells deficient in proteinfucosylation (Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986);U.S. Pat Appl No 2003/0157108 A1, Presta, L; and WO 2004/056312 A1,Adams et al., especially at Example 11), and knockout cell lines, suchas alpha-1,6-fucosyltransferase gene, FUT8, knockout CHO cells (see,e.g., Yamane-Ohnuki et al. Biotech. Bioeng. 87: 614 (2004); Kanda, Y. etal., Biotechnol. Bioeng., 94(4):680-688 (2006); and WO2003/085107).

Antibodies variants are further provided with bisected oligosaccharides,e.g., in which a biantennary oligosaccharide attached to the Fc regionof the antibody is bisected by GlcNAc. Such antibody variants may havereduced fucosylation and/or improved ADCC function. Examples of suchantibody variants are described, e.g., in WO 2003/011878 (Jean-Mairet etal.); U.S. Pat. No. 6,602,684 (Umana et al.); and US 2005/0123546 (Umanaet al.). Antibody variants with at least one galactose residue in theoligosaccharide attached to the Fc region are also provided. Suchantibody variants may have improved CDC function. Such antibody variantsare described, e.g., in WO 1997/30087 (Patel et al.); WO 1998/58964(Raju, S.); and WO 1999/22764 (Raju, S.).

Fc Region Variants

In certain embodiments, one or more amino acid modifications may beintroduced into the Fc region of an antibody provided herein, therebygenerating an Fc region variant. The Fc region variant may comprise ahuman Fc region sequence (e.g., a human IgG1, IgG2, IgG3 or IgG4 Fcregion) comprising an amino acid modification (e.g. a substitution) atone or more amino acid positions.

In certain embodiments, the invention contemplates an antibody variantthat possesses some but not all effector functions, which make it adesirable candidate for applications in which the half-life of theantibody in vivo is important yet certain effector functions (such ascomplement and ADCC) are unnecessary or deleterious. In vitro and/or invivo cytotoxicity assays can be conducted to confirm thereduction/depletion of CDC and/or ADCC activities. For example, Fcreceptor (FcR) binding assays can be conducted to ensure that theantibody lacks FcyR binding (hence likely lacking ADCC activity), butretains FcRn binding ability. The primary cells for mediating ADCC, NKcells, express FcyRIII only, whereas monocytes express FcyRI, FcyRII andFcyRIII FcR expression on hematopoietic cells is summarized in Table 3on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9:457-492 (1991).Non-limiting examples of in vitro assays to assess ADCC activity of amolecule of interest is described in U.S. Pat. No. 5,500,362 (see, e.g.Hellstrom, I. et al. Proc. Nat'l Acad. Sci. USA 83:7059-7063 (1986)) andHellstrom, I et al., Proc. Nat'l Acad. Sci. USA 82:1499-1502 (1985);5,821,337 (see Bruggemann, M. et al., J. Exp. Med. 166:1351-1361(1987)). Alternatively, non-radioactive assays methods may be employed(see, for example, ACTITM non-radioactive cytotoxicity assay for flowcytometry (CellTechnology, Inc. Mountain View, Calif.; and CytoTox 96®non-radioactive cytotoxicity assay (Promega, Madison, Wis.). Usefuleffector cells for such assays include peripheral blood mononuclearcells (PBMC) and Natural Killer (NK) cells. Alternatively, oradditionally, ADCC activity of the molecule of interest may be assessedin vivo, e.g., in an animal model such as that disclosed in Clynes etal. Proc. Nat'l Acad. Sci. USA 95:652-656 (1998). C1q binding assays mayalso be carried out to confirm that the antibody is unable to bind C1qand hence lacks CDC activity. See, e.g., C1q and C3c binding ELISA in WO2006/029879 and WO 2005/100402. To assess complement activation, a CDCassay may be performed (see, for example, Gazzano-Santoro et al., J.Immunol. Methods 202:163 (1996); Cragg, M. S. et al., Blood101:1045-1052 (2003); and Cragg, M. S. and M. J. Glennie, Blood103:2738-2743 (2004)). FcRn binding and in vivo clearance/half-lifedeterminations can also be performed using methods known in the art(see, e.g., Petkova, S. B. et al., Int'l. Immunol. 18(12):1759-1769(2006)).

Antibodies with reduced effector function include those withsubstitution of one or more of Fc region residues 238, 265, 269, 270,297, 327 and 329 (U.S. Pat. No. 6,737,056). Such Fc mutants include Fcmutants with substitutions at two or more of amino acid positions 265,269, 270, 297 and 327, including the so-called “DANA” Fc mutant withsubstitution of residues 265 and 297 to alanine (U.S. Pat. No.7,332,581).

Certain antibody variants with improved or diminished binding to FcRsare described. (See, e.g., U.S. Pat. No. 6,737,056; WO 2004/056312, andShields et al., J. Biol. Chem. 9(2): 6591-6604 (2001).)

In certain embodiments, an antibody variant comprises an Fc region withone or more amino acid substitutions which improve ADCC, e.g.,substitutions at positions 298, 333, and/or 334 of the Fc region (EUnumbering of residues).

In some embodiments, alterations are made in the Fc region that resultin altered (i.e., either improved or diminished) C1q binding and/orComplement Dependent Cytotoxicity (CDC), e.g., as described in U.S. Pat.No. 6,194,551, WO 99/51642, and Idusogie et al. J. Immunol. 164:4178-4184 (2000).

Antibodies with increased half-lives and improved binding to theneonatal Fc receptor (FcRn), which is responsible for the transfer ofmaternal IgGs to the fetus (Guyer et al., J. Immunol. 117:587 (1976) andKim et al., J. Immunol. 24:249 (1994)), are described inUS2005/0014934A1 (Hinton et al.). Those antibodies comprise an Fc regionwith one or more substitutions therein which improve binding of the Fcregion to FcRn. Such Fc variants include those with substitutions at oneor more of Fc region residues: 238, 256, 265, 272, 286, 303, 305, 307,311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or 434,e.g., substitution of Fc region residue 434 (U.S. Pat. No. 7,371,826).

See also Duncan & Winter, Nature 322:738-40 (1988); U.S. Pat. No.5,648,260; U.S. Pat. No. 5,624,821; and WO 94/29351 concerning otherexamples of Fc region variants.

Cysteine Engineered Antibody Variants

In certain embodiments, it may be desirable to create cysteineengineered antibodies, e.g., “THIOMABS,” in which one or more residuesof an antibody are substituted with cysteine residues. In particularembodiments, the substituted residues occur at accessible sites of theantibody. By substituting those residues with cysteine, reactive thiolgroups are thereby positioned at accessible sites of the antibody andmay be used to conjugate the antibody to other moieties, such as drugmoieties or linker-drug moieties, to create an immunoconjugate, asdescribed further herein. In certain embodiments, any one or more of thefollowing residues may be substituted with cysteine: V205 (Kabatnumbering) of the light chain; A118 (EU numbering) of the heavy chain;and S400 (EU numbering) of the heavy chain Fc region. Cysteineengineered antibodies may be generated as described, e.g., in U.S. Pat.No. 7,521,541.

Antibody Derivatives

In certain embodiments, an antibody provided herein may be furthermodified to contain additional nonproteinaceous moieties that are knownin the art and readily available. The moieties suitable forderivatization of the antibody include but are not limited to watersoluble polymers. Non-limiting examples of water soluble polymersinclude, but are not limited to, polyethylene glycol (PEG), copolymersof ethylene glycol/propylene glycol, carboxymethylcellulose, dextran,polyvinyl alcohol, polyvinyl pyrrolidone, poly-1, 3-dioxolane,poly-1,3,6-trioxane, ethylene/maleic anhydride copolymer, polyaminoacids(either homopolymers or random copolymers), and dextran or poly(n-vinylpyrrolidone)polyethylene glycol, propropylene glycol homopolymers,polypropylene oxide/ethylene oxide co-polymers, polyoxyethylated polyols(e.g., glycerol), polyvinyl alcohol, and mixtures thereof. Polyethyleneglycol propionaldehyde may have advantages in manufacturing due to itsstability in water. The polymer may be of any molecular weight, and maybe branched or unbranched. The number of polymers attached to theantibody may vary, and if more than one polymer is attached, they can bethe same or different molecules. In general, the number and/or type ofpolymers used for derivatization can be determined based onconsiderations including, but not limited to, the particular propertiesor functions of the antibody to be improved, whether the antibodyderivative will be used in a therapy under defined conditions, etc.

In another embodiment, conjugates of an antibody and nonproteinaceousmoiety that may be selectively heated by exposure to radiation areprovided. In one embodiment, the nonproteinaceous moiety is a carbonnanotube (Kam et al., Proc. Natl. Acad. Sci. USA 102: 11600-11605(2005)). The radiation may be of any wavelength, and includes, but isnot limited to, wavelengths that do not harm ordinary cells, but whichheat the nonproteinaceous moiety to a temperature at which cellsproximal to the antibody-nonproteinaceous moiety are killed.

Recombinant Methods and Compositions

Antibodies may be produced using recombinant methods and compositions,e.g., as described in U.S. Pat. No. 4,816,567. In one embodiment,isolated nucleic acid encoding an antibody described herein is provided.Such nucleic acid may encode an amino acid sequence comprising the VLand/or an amino acid sequence comprising the VH of the antibody (e.g.,the light and/or heavy chains of the antibody). In a further embodiment,one or more vectors (e.g., expression vectors) comprising such nucleicacid are provided. In a further embodiment, a host cell comprising suchnucleic acid is provided. In one such embodiment, a host cell comprises(e.g., has been transformed with): (1) a vector comprising a nucleicacid that encodes an amino acid sequence comprising the VL of theantibody and an amino acid sequence comprising the VH of the antibody,or (2) a first vector comprising a nucleic acid that encodes an aminoacid sequence comprising the VL of the antibody and a second vectorcomprising a nucleic acid that encodes an amino acid sequence comprisingthe VH of the antibody. In one embodiment, the host cell is eukaryotic,e.g. a Chinese Hamster Ovary (CHO) cell or lymphoid cell (e.g., YO, NSO,Sp20 cell). In one embodiment, a method of making an antibody isprovided, wherein the method comprises culturing a host cell comprisinga nucleic acid encoding the antibody, as provided above, underconditions suitable for expression of the antibody, and optionallyrecovering the antibody from the host cell (or host cell culturemedium).

For recombinant production of an antibody, nucleic acid encoding anantibody, e.g., as described above, is isolated and inserted into one ormore vectors for further cloning and/or expression in a host cell. Suchnucleic acid may be readily isolated and sequenced using conventionalprocedures (e.g., by using oligonucleotide probes that are capable ofbinding specifically to genes encoding the heavy and light chains of theantibody).

Suitable host cells for cloning or expression of antibody-encodingvectors include prokaryotic or eukaryotic cells described herein. Forexample, antibodies may be produced in bacteria, in particular whenglycosylation and Fc effector function are not needed. For expression ofantibody fragments and polypeptides in bacteria, see, e.g., U.S. Pat.Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods inMolecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, N.J.,2003), pp. 245-254, describing expression of antibody fragments in E.coli.) After expression, the antibody may be isolated from the bacterialcell paste in a soluble fraction and can be further purified.

In addition to prokaryotes, eukaryotic microbes such as filamentousfungi or yeast are suitable cloning or expression hosts forantibody-encoding vectors, including fungi and yeast strains whoseglycosylation pathways have been “humanized,” resulting in theproduction of an antibody with a partially or fully human glycosylationpattern. See Gerngross, Nat. Biotech. 22:1409-1414 (2004), and Li etal., Nat. Biotech. 24:210-215 (2006).

Suitable host cells for the expression of glycosylated antibody are alsoderived from multicellular organisms (invertebrates and vertebrates).Examples of invertebrate cells include plant and insect cells. Numerousbaculoviral strains have been identified which may be used inconjunction with insect cells, particularly for transfection ofSpodoptera frugiperda cells.

Plant cell cultures can also be utilized as hosts. See, e.g., U.S. Pat.Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429(describing PLANTIBODIESTM technology for producing antibodies intransgenic plants).

Vertebrate cells may also be used as hosts. For example, mammalian celllines that are adapted to grow in suspension may be useful. Otherexamples of useful mammalian host cell lines are monkey kidney CV1 linetransformed by SV40 (COS-7); human embryonic kidney line (293 or 293cells as described, e.g., in Graham et al., J. Gen Virol. 36:59 (1977));baby hamster kidney cells (BHK); mouse sertoli cells (TM4 cells asdescribed, e.g., in Mather, Biol. Reprod. 23:243-251 (1980)); monkeykidney cells (CV1); African green monkey kidney cells (VERO-76); humancervical carcinoma cells (HELA); canine kidney cells (MDCK; buffalo ratliver cells (BRL 3A); human lung cells (W138); human liver cells (HepG2); mouse mammary tumor (MMT 060562); TRI cells, as described, e.g., inMather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982); MRC 5 cells; andFS4 cells. Other useful mammalian host cell lines include Chinesehamster ovary (CHO) cells, including DHFR-CHO cells (Urlaub et al.,Proc. Natl. Acad. Sci. USA 77:4216 (1980)); and myeloma cell lines suchas YO, NSO and Sp2/0. For a review of certain mammalian host cell linessuitable for antibody production, see, e.g., Yazaki and Wu, Methods inMolecular Biology, Vol. 248 (B. K. C. Lo, ed., Humana Press, Totowa,N.J.), pp. 255-268 (2003).

Pharmaceutical Formulations

Pharmaceutical formulations, also referred to herein as pharmaceuticalcompositions, of an anti-IL-13 antibody as described herein are preparedby mixing such antibody or molecule having the desired degree of puritywith one or more optional pharmaceutically acceptable carriers(Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)),in the form of lyophilized formulations or aqueous solutions.Pharmaceutically acceptable carriers are generally nontoxic torecipients at the dosages and concentrations employed, and include, butare not limited to: buffers such as phosphate, citrate, and otherorganic acids; antioxidants including ascorbic acid and methionine;preservatives (such as octadecyldimethylbenzyl ammonium chloride;hexamethonium chloride; benzalkonium chloride; benzethonium chloride;phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol);low molecular weight (less than about 10 residues) polypeptides;proteins, such as serum albumin, gelatin, or immunoglobulins;hydrophilic polymers such as polyvinylpyrrolidone; amino acids such asglycine, glutamine, asparagine, histidine, arginine, or lysine;monosaccharides, disaccharides, and other carbohydrates includingglucose, mannose, or dextrins; chelating agents such as EDTA; sugarssuch as sucrose, mannitol, trehalose or sorbitol; salt-formingcounter-ions such as sodium; metal complexes (e.g. Zn-proteincomplexes); and/or non-ionic surfactants such as polyethylene glycol(PEG). Exemplary pharmaceutically acceptable carriers herein furtherinclude insterstitial drug dispersion agents such as solubleneutral-active hyaluronidase glycoproteins (sHASEGP), for example, humansoluble PH-20 hyaluronidase glycoproteins, such as rHuPH20 (HYLENEX®,Baxter International, Inc.). Certain exemplary sHASEGPs and methods ofuse, including rHuPH20, are described in US Patent Publication Nos.2005/0260186 and 2006/0104968. In one aspect, a sHASEGP is combined withone or more additional glycosaminoglycanases such as chondroitinases.

Exemplary lyophilized antibody formulations are described in U.S. Pat.No. 6,267,958. Aqueous antibody formulations include those described inU.S. Pat. No. 6,171,586 and WO2006/044908, the latter formulationsincluding a histidine-acetate buffer.

The formulation herein may also contain more than one active ingredientsas necessary for the particular indication being treated, preferablythose with complementary activities that do not adversely affect eachother. For example, it may be desirable to further provide a controllerwith the Th2 pathway inhibitor. Such active ingredients are suitablypresent in combination in amounts that are effective for the purposeintended.

Active ingredients may be entrapped in microcapsules prepared, forexample, by coacervation techniques or by interfacial polymerization,for example, hydroxymethylcellulose or gelatin-microcapsules andpoly-(methylmethacylate) microcapsules, respectively, in colloidal drugdelivery systems (for example, liposomes, albumin microspheres,microemulsions, nano-particles and nanocapsules) or in macroemulsions.Such techniques are disclosed in Remington's Pharmaceutical Sciences16th edition, Osol, A. Ed. (1980).

Sustained-release preparations may be prepared. Suitable examples ofsustained-release preparations include semipermeable matrices of solidhydrophobic polymers containing the antibody, which matrices are in theform of shaped articles, e.g. films, or microcapsules.

The formulations to be used for in vivo administration are generallysterile. Sterility may be readily accomplished, e.g., by filtrationthrough sterile filtration membranes.

Therapeutic Methods and Compositions

In certain embodiments, methods of treating atopic dermatitis comprisingadministering a therapeutically effective amount of lebrikizumab to theatopic dermatitis patient, wherein the treatment results in a reductionin disease severity as measured or assessed by an Atopic DermatitisDisease Severity Outcome Measure (ADDSOM) are provided. In someembodiments, the atopic dermatitis is moderate to severe as determinedby Rajka/Langeland criteria score. A Rajka/Langeland criteria scorebetween 4.5 and 9 is typically considered moderate to severe atopicdermatitis. In some embodiments, the methods further compriseadministration of one or more topical corticosteroids. Topicalcorticosteroids may be administered before (i.e., prior to)administration of the IL-13 antagonist, at the same time asadministration of the IL-13 antagonist, or after administration of theIL-13 antagonist. Exemplary topical corticosteroids include, but are notlimited to, triamcinolone acetonide, hydrocortisone, and a combinationof triamcinolone acetonide and hydrocortisone. Triamcinolone acetonideis typically formulated at a concentration of 0.1% in a cream, andhydrocortisone is typically formulated at a concentration of 1% or 2.5%in a cream. Certain topical corticosteroids are considered very highpotency such as, for example, betamethasone dipropionate, clobetasolpropionate, diflorasone diacetate, fluocinonide, and halobetasolpropionate. Certain topical corticosteroids are considered high potencysuch as, for example, amcinonide, desoximetasone, halcinonide, andtriamcinolone acetonide. Certain topical corticosteroids are consideredmedium potency, such as, for example, betamethasone valerate,clocortolone pivalate, fluocinolone acetonide, flurandrenolide,fluocinonide, fluticasone propionate, hydrocortisone butyrate,hydrocortisone valerate, mometasone furoate, and prednicarbate. Certaintopical corticosteroids are considered low potency, such as, forexample, alclometasone dipropionate, desonide, and hydrocortisone. TCSmay be applied to affected areas once daily, twice daily, three timesper day, or as needed. In some embodiments, the patient is inadequatelycontrolled on topical corticosteroids.

Various ADDSOMs are known in the art and are described herein inaddition to a novel Atopic Dermatitis Impact Questionnaire (ADIQ).Exemplary ADDSOMs include, but are not limited to, “Eczema Area andSeverity Index” (EAST), “Severity Scoring of Atopic Dermatitis”(SCORAD), “Investigator Global Assessment” (IGA), “Investigator GlobalAssessment of Signs” (IGSA), Rajka/Langeland Atopic Dermatitis SeverityScore, and Patient-Reported Outcomes including, but not limited to,Pruritus Visual Analog Scale (an aspect of disease severity assessed aspart of SCORAD), Sleep Loss Visual Analog Scale (an aspect of diseaseseverity assessed as part of SCORAD), Atopic Dermatitis Symptom Diary(ADSD), Atopic Dermatitis Impact Questionnaire (ADIQ), Dermatology LifeQuality Index (DLQI) (Finlay and Khan, Clin Exper Dermatol 1994;19:210),and 5-D Itch Scale (Elman et al., Br J Dermatol 2010;162(3):587-593).

A therapeutic agent as provided herein can be administered by anysuitable means, including parenteral, subcutaneous, intraperitoneal,intrapulmonary, and intranasal. Parenteral infusions includeintramuscular, intravenous, intraarterial, intraperitoneal, orsubcutaneous administration. In certain embodiments, the dosing is givenby injections, e.g., intravenous or subcutaneous injections. In yetanother embodiment, the therapeutic agent is administered using asyringe (e.g., prefilled or not) or an autoinjector. In certainembodiments, the therapeutic agent is applied topically.

In certain embodiments, an anti-IL13 antibody of the invention isadministered using, for example, a self-inject device, autoinjectordevice, or other device designed for self-administration. In certainembodiments, an anti-IL13 antibody of the invention is administeredusing a subcutaneous administration device. Various self-inject devicesand subcutaneous administration devices, including autoinjector devices,are known in the art and are commercially available. Exemplary devicesinclude, but are not limited to, prefilled syringes (such as BD HYPAKSCF®, READYFILL™, and STERIFILL SCF™ from Becton Dickinson; CLEARSHOT™copolymer prefilled syringes from Baxter; and Daikyo Seiko CRYSTALZENITH® prefilled syringes available from West Pharmaceutical Services);disposable pen injection devices such as BD Pen from Becton Dickinson;ultra-sharp and microneedle devices (such as INJECT-EASE™ andmicroinfuser devices from Becton Dickinson; and H-PATCH™ available fromValeritas) as well as needle-free injection devices (such as BIOJECTOR®and IJECT® available from Bioject; and SOF-SERTER® and patch devicesavailable from Medtronic). In certain embodiments, an anti-IL-13antibody of the invention is administered using an autoinjector devicecomprised of various components, for example, as described in US PatentPublication Nos. 2014/0114247, 2014/0148763, and 2013/0131590; U.S. Pat.Nos. 7,597,685, 7,896,850, and 8,617,109, all of which are incorporatedby reference. Co-formulations or co-administrations with suchself-inject devices or subcutaneous administration devices of ananti-IL13 antibody with at least a second therapeutic compound areenvisioned

For the prevention or treatment of disease, the appropriate dosage of anantibody of the invention (when used alone or in combination with one ormore other additional therapeutic agents) will depend on the type ofdisease to be treated, the type of antibody, the severity and course ofthe disease, whether the antibody is administered for preventive ortherapeutic purposes, previous therapy, the patient's clinical historyand response to the antibody, and the discretion of the attendingphysician. The antibody is suitably administered to the patient at onetime or over a series of treatments. In certain embodiments, theantibody of the invention is administered as a flat dose (not aweight-based dose) of 125 mg or 250 mg or about 125 mg or about 250 mgor a flat dose between 110 mg to 140 mg or a flat dose between 120 mg to130 mg or a flat dose between 225 mg to 275 mg or a flat dose between240 mg to 260 mg. In certain embodiments, the dose is administered bysubcutaneous injection once every four weeks or once every eight weeksfor a period of time. In certain embodiments, the period of time is 12weeks, or 20 weeks, or 24 weeks, or 9 months, or one year, two years,five years, ten years, 15 years, 20 years, or the lifetime of thepatient. The progress of this therapy is easily monitored byconventional techniques and assays. In certain embodiments, the patienthas moderate to severe atopic dermatitis and the patient administers TCSas needed while being treated with the antibody of the invention. Incertain embodiments, the TCS is administered once a day, or twice a day,or three times a day or more. In certain embodiments, the patient isable to decrease usage of TCS over time while the antibody of theinvention is being administered. Such decreased usage of TCS is referredto as “tapering” or “TCS-sparing.” The time period over which decreasedusage may occur is one, two, three, or four weeks, or two, three, four,five, or six months.

IL-13 antagonists of the invention, according to certain embodiments,comprise administration in combination with one or more additionaltherapeutic agents. The term “in combination with” means that theadditional therapeutic agent(s) are administered before, after, orconcurrent with the pharmaceutical composition comprising the IL-13antagonist. The term “in combination with” also includes sequential orconcomitant administration of IL-13 antagonist and a second therapeuticagent.

For example, when administered “before” the pharmaceutical compositioncomprising the IL-13 antagonist, the additional therapeutic agent may beadministered about 72 hours, about 60 hours, about 48 hours, about 36hours, about 24 hours, about 12 hours, about 10 hours, about 8 hours,about 6 hours, about 4 hours, about 2 hours, about 1 hour, about 30minutes, about 15 minutes or about 10 minutes prior to theadministration of the pharmaceutical composition comprising the IL-13antagonist. When administered “after” the pharmaceutical compositioncomprising the IL-13 antagonist, the additional therapeutic agent may beadministered about 10 minutes, about 15 minutes, about 30 minutes, about1 hour, about 2 hours, about 4 hours, about 6 hours, about 8 hours,about 10 hours, about 12 hours, about 24 hours, about 36 hours, about 48hours, about 60 hours or about 72 hours after the administration of thepharmaceutical composition comprising the IL-13 antagonist.Administration “concurrent” or with the pharmaceutical compositioncomprising the IL-13 antagonist means that the additional therapeuticagent is administered to the subject in a separate dosage form withinless than 5 minutes (before, after, or at the same time) ofadministration of the pharmaceutical composition comprising the IL-13antagonist, or administered to the subject as a single combined dosageformulation comprising both the additional therapeutic agent and theIL-13 antagonist.

The additional therapeutic agent may be, e.g., another IL-13 antagonist,an IL-4R antagonist, an IL-1 antagonist, an IL-6 antagonist, an IL-6Rantagonist, a TNF antagonist, an IL-8 antagonist, an IL-9 antagonist, anIL-17 antagonist, an IL-5 antagonist, an IgE antagonist, a CD48antagonist, an IL-31 antagonist, a thymic stromal lymphopoietin (TSLP)antagonist, interferon-gamma (IFN.gamma.) antibiotics, topicalcorticosteroids, tacrolimus, pimecrolimus, mycophenolate, cyclosporine,azathioprine, methotrexate, cromolyn sodium, proteinase inhibitors, orcombinations thereof In certain embodiments, the pharmaceuticalcomposition comprising an anti-IL13 antagonist is administered to asubject in conjunction with an oral immunosuppressant, a topicalcalcineurin inhibitor, such as for example, pimecrolimus or tacrolimus,or an oral corticosteroid, such as, for example, prednisolone. Incertain embodiments, the pharmaceutical composition comprising ananti-IL13 antagonist is administered to a subject in conjunction with anon-pharmaceutical therapy such as ultraviolet (UV) light therapy.

In certain embodiments, an antibody of the invention is administered oneor more times as a flat (i.e., not weight dependent) loading dosefollowed by administration one or more times as a flat maintenance dose.In certain embodiments, the loading dose is 250 mg or 500 mg and themaintenance dose 125 mg. In certain embodiments, two loading doses of250 mg are administered 15 days apart or 29 days apart. In certainembodiments, the maintenance dose is administered two weeks after thelast loading dose or four weeks after the last loading dose and thenevery four weeks thereafter. In certain embodiments, the loading dose is500 mg and the maintenance dose is 250 mg. In certain embodiments, themaintenance dose of 250 mg is administered four weeks after the loadingdose and then every four weeks or every eight weeks thereafter.

Articles of Manufacture

In another aspect of the invention, an article of manufacture containingmaterials useful for the treatment, prevention and/or diagnosis of thedisorders described above is provided. The article of manufacturecomprises a container and a label or package insert on or associatedwith the container. Suitable containers include, for example, bottles,vials, syringes, IV solution bags, autoinjectors etc. The containers maybe formed from a variety of materials such as glass or plastic. Thecontainer holds a composition which is by itself or combined withanother composition effective for treating, preventing and/or diagnosingthe condition and may have a sterile access port (for example thecontainer may be an intravenous solution bag or a vial having a stopperpierceable by a hypodermic injection needle). At least one active agentin the composition is an antibody of the invention. The label or packageinsert indicates that the composition is used for treating the conditionof choice. Moreover, the article of manufacture may comprise (a) a firstcontainer with a composition contained therein, wherein the compositioncomprises an antibody of the invention; and (b) a second container witha composition contained therein, wherein the composition comprises afurther therapeutic agent. The article of manufacture in this embodimentof the invention may further comprise a package insert indicating thatthe compositions can be used to treat a particular condition.Alternatively, or additionally, the article of manufacture may furthercomprise a second (or third) container comprising apharmaceutically-acceptable buffer, such as bacteriostatic water forinjection (BWFI), phosphate-buffered saline, Ringer's solution anddextrose solution. It may further include other materials desirable froma commercial and user standpoint, including other buffers, diluents,filters, needles, syringes, and autoinjectors.

EXAMPLES Example 1 Anti-IL-13 Antibodies

Anti-IL-13 antibodies, including monoclonal humanized anti-IL-13antibodies, have been described previously (see, e.g., WO2005062967).One monoclonal humanized anti-IL-13 antibody described therein islebrikizumab, an IgG4 antibody, see also Sequence Listing Table.Lebrikizumab was used in the studies described below, formulated at 125mg/mL in a pharmaceutically acceptable composition and supplied in aprefilled syringe.

Example 2 Clinical Study I

Clinical Study I was a phase II, randomized, double-blind,placebo-controlled study to evaluate the safety and efficacy oflebrikizumab in patients (age 18-75) with persistent moderate to severeatopic dermatitis that is inadequately controlled by topicalcorticosteroids (TCS). In this Study, lebrikizumab was tested incombination with TCS. The Study Schema is provided in FIG. 1.

Screening. Patients eligible to enroll in the Study had to have moderateto severe AD present for at least 1 year and had to meet all eligibilitycriteria. Patients were evaluated at screening to assess eligibility forentry into the run-in period and again at the end of the run-in period,to assess eligibility for entry into the treatment period andrandomization to study drug.

The inclusion criteria included the following: age 18 to 75 years; ADdiagnosed by the Hanifin/Rajka criteria and that had been present for atleast 1 year at screening; moderate to severe AD as graded by theRajka/Langeland criteria at screening; history of inadequate response toa 1 month (within the 3 months prior to the screening visit) treatmentregimen of at least daily TCS and regular emollient for treatment of AD;EASI score 14 at screening and end of the run-in period (Visit 3), IGAscore 3 (5-point scale) at screening and end of the run-in period (Visit3); AD involvement of 10% body surface area (BSA) at screening; andPruritus VAS score (measured as part of the SCORAD) of 3 at screening.

Exclusion criteria included the following: past and/or current use ofany anti-IL-13 or anti-IL-4/IL-13 therapy, including lebrikizumab; useof an investigational agent within 4 weeks prior to screening or within5 half-lives of the investigational agent, whichever is longer; historyof a severe allergic reaction or anaphylactic reaction to a biologicagent or known hypersensitivity to any component of the lebrikizumabinjection; hypersensitivity to TCS or to any other ingredients containedby the TCS product used in the study; use of any complementary,alternative, or homeopathic medicines including, but not limited to,phytotherapies, traditional or non-traditional herbal medications,essential fatty acids, or acupuncture within 7 days prior to the run-inperiod or need for such medications during the study; body weight <40 kgor body mass index >38 kg/m²; evidence of other skin conditions;including, but not limited to, T-cell lymphoma or allergic contactdermatitis; evidence of, or ongoing treatment (including topicalantibiotics) for active skin infection at screening (Day −15); certaininfections; history of recent (<1 year) parasitic infections, especiallynematodes (e.g., Ascaris, Ancylostoma), Platyhelminthes (e.g.,Schistosoma), or history of Listeria infections; active tuberculosisrequiring treatment within the 12 months prior to Visit 1; evidence ofacute or chronic hepatitis or known liver cirrhosis; knownimmunodeficiency, including HIV infection; use of TCI (topicalcalcineurin inhibitor) at the time of screening; use of a tanningbooth/parlor within 4 weeks before the screening visit; allergenimmunotherapy within 3 months of screening; receipt of a live attenuatedvaccine within 4 weeks prior to baseline visit (Day 1); planned surgeryduring the study; clinically significant abnormality on screening ECG orlaboratory tests (hematology, serum chemistry, and urinalysis); AST,ALT, or total bilirubin elevation ≥2.0×the upper limit of normal (ULN)during screening; known current malignancy or current evaluation for apotential malignancy, including basal or squamous cell carcinoma of theskin or carcinoma in situ; history of malignancy within 5 years prior toscreening, except for appropriately treated carcinoma in situ of thecervix, non-melanoma skin carcinoma; stage I uterine cancer; and otherclinically significant medical disease that is uncontrolled despitetreatment.

Run-in Period. Following the screening visit, eligible patients entereda 2-week run-in period (Days −14 to −1) during which aprotocol-specified topical therapy regimen was initiated. On the firstday of the run-in period (Day −14) patients received TCS for usethroughout the run-in period. Patients were required to apply thefollowing topical therapy regimen daily: emollient to all xerotic skinsurfaces, at least once daily; TCS cream consisting of medium potencyTCS (triamcinolone acetonide 0.1%), two times per day, only to allactive skin lesions. For lesions affecting the face or intertriginousareas, low potency TCS (hydrocortisone 2.5% cream) could be used insteadof triamcinolone acetonide 0.1%, at the discretion of the investigator.

At the end of the run-in period, a second assessment of eligibility forrandomization to study drug (lebrikizumab or placebo) was performed andpatients must have continued to demonstrate moderate to severe AD asassessed by EASI score ≥14 and IGA score ≥3.

Treatment Period. At the end of the run-in period, patients who had: 1)demonstrated compliance with the protocol-specified TCS regimen, and 2)who continued to fulfill the eligibility criteria as described abovewere randomized. The plan was to randomize approximately 200 patients(1:1:1:1) to one of the following four treatment groups (see FIG. 1):(1) Group 1: lebrikizumab 250 mg administered subcutaneously (SC) singledose (Day 1) followed by 2 placebo doses (at Week 4 and at Week 8) for atotal of 3 doses+TCS cream; (2) Group 2: lebrikizumab 125 mg SC singledose (Day 1) followed by 2 placebo doses (Week 4 and Week 8) for a totalof 3 doses+TCS cream; (3) Group 3: lebrikizumab 125 mg SC every 4 weeks(Q4W) for a total of 3 doses+TCS cream; and (4) Group 4: placebo SC Q4Wfor a total of 3 doses+TCS cream. Three active doses were included tocharacterize both exposure-response relationships and dosing frequencyrequirements.

Patients randomized to these four study treatment groups received studydrug (lebrikizumab or placebo SC) in combination with TCS. Throughoutthe 12-week treatment period these patients continued to apply emollientat least once daily and triamcinolone acetonide 0.1% cream as describedin the run-in period (two times per day to all active skin lesions onthe body). For lesions affecting the face or intertriginous areas, lowpotency TCS (hydrocortisone 2.5% cream) could have been used instead oftriamcinolone acetonide 0.1%, at the discretion of the investigator.Patients recorded their use of topical creams and responded toPRO-related questions using a hand held eDiary throughout the treatmentperiod. At each study visit the patient was required to bring their TCScontainers to the study site to be weighed.

Safety Follow-Up Period. All patients who completed study treatmentduring the placebo-controlled treatment period (Weeks 1-12) werefollowed for safety for an additional 8 weeks (Weeks 13-20). During thissafety follow-up period, patients were no longer required to apply thetopical treatment regimen as specified during the run-in and treatmentperiods. Instead, triamcinolone acetonide 0.1% cream and/orhydrocortisone 2.5% cream could be applied as determined by the patientand the study investigator. At each study visit the patient was requiredto bring their TCS containers to the study site to be weighed. Patientswere encouraged to continue applying emollients at least once daily toxerotic skin. An escalation in therapy for AD was allowed at any duringthe study, if, in the opinion of the investigator, it was clinicallyindicated.

Topical calcineruin inhibitors (TCIs) were not allowed at any timeduring this study. Patients who had been using TCIs at the time ofscreening were allowed to participate in the study if they agreed tostop using TCIs during the study and if, in the opinion of theinvestigator, it was safe to stop the use of TCI and use the protocolspecified TCS cream instead.

Rationale for Lebrikizumab Dose and Schedule. The dose of lebrikizumabfor AD was based on the clinical experience with the asthma program andthe similarities in IL-13 biology and the expected PK characteristics oflebrikizumab in the two patient populations. Both AD and asthma patientshave elevated levels of biomarkers associated with IL-13 biology andTh-2 mediated hypersensitivity. In particular, IL-13 is elevated in thelungs of asthma patients and has a pathogenic role in asthma, whileincreased expression of IL-13 has consistently been reported in AD skin.

Additionally, given that the primary clearance mechanism of lebrikizumabis through nonspecific endocytosis and catabolism, with minimalcontribution of target-mediated clearance, the PK characteristics oflebrikizumab are expected to be similar between asthma and AD patients.Although limited information exists in the literature regarding thepartitioning of monoclonal antibodies to various tissues, available datain animal models suggest similar plasma-to-tissue partitioningcoefficients for monoclonal antibodies in the lung and skin tissue (ShahD K et al., mAbs 2013; 5(2):297-305). Therefore, the dose levels oflebrikizumab producing clinical benefit in asthma may also producebiological activity in AD patients.

As described above, three different dose regimens were tested in thisPhase II study: (1) the 125 mg Q4W dose (Group 3) was included as apotentially effective dose. 125-mg Q4W was the highest dose regimen(i.e. highest total exposure) being studied in the pivotal Phase IIIadult studies of lebrikizumab in asthma (LAVOLTA I and II) that wereongoing at the time of study initiation. Given the expected similarityin the role of IL-13 and the pharmacokinetics between asthma and AD, itwas hypothesized that this dose could be effective in patients with AD;(2) a single 250 mg dose (Group 1) was proposed as an alternativepotentially effective dose. This dose was expected to produce a loweroverall exposure than 125 mg Q4W over the first 12 weeks. With theprimary endpoint at Week 12, a single 250 mg dose was tested to explorethe potential for quarterly dosing; and (3) a single 125 mg dose (Group2) was proposed as a potentially partially effective dose. Given thatthe overall exposure at this dose was expected to be 2-3-fold lower thanthat of the 125 mg Q4W and 250 mg single dose regimens, data from allthree dose arms can be used to characterize the exposure-responserelationships of lebrikizumab in AD patients.

As described above, however, the results of the LAVOLTA I and II studieswere inconsistent. Hanania et al., Lancet Respir Med 2016, available atdx(dot)doi(dot)org(slash)52213-2600(16)30265-X, published onlineSeptember 5, 2016. Both studies failed to show a clear dose-responsealthough pharmacokinetic and pharmacodynamics results were consistentwith those from the phase II studies described previously, indicatingthat drug exposures were similar and that the IL-13 pathway wasinhibited. Hanania et al., 2016, supra. Accordingly, it was uncertainwhether lebrikizumab would provide clinically meaningful benefit toatopic dermatitis patients at the dosing regimens described herein.

The primary endpoint for the Phase II study was measured at Week 12. Themean serum lebrikizumab trough concentration at Week 12 was expected tobe over 90% of the steady state value for patients receiving 125 mg Q4W.In addition, for the 125 mg Q4W dose, the predicted troughconcentrations at Week 12 would be well within the range wherelebrikizumab demonstrated biological activity in the Phase II asthmaclinical trials. For the 250 mg single dose, the predicted averageexposure over 12 weeks is above that of a 37.5 mg Q4W regimen, whichalso demonstrated biological activity in Phase II asthma trials; Hananiaet al., Thorax 2015;70:748-756.

Rationale for Control Group. Patients in treatment Group 4 receivedplacebo SC+TCS cream and therefore served as an active comparator. Thistreatment group was used as a control for lebrikizumab SC to adequatelyassess the efficacy and safety effect of lebrikizumab as adjunctivetherapy with TCS.

The active TCS comparator group was included since TCS use is thestandard of care in clinical practice and is continued while step-uptherapies (e.g., systemic agents) are initiated (European Academy ofDermatology and Venereology [EADV] 2009 guidelines; Darsow et al., J EurAcad Dermatol Venereol 2010; 24:317-28). In addition, stopping TCS inpatients with persistent moderate to severe disease may result insignificant worsening of AD symptoms and disease burden.

Efficacy Outcome Measures. The primary efficacy outcome measure was thepercentage of patients achieving EASI-50 (a 50% reduction in EASI scorefrom baseline) at Week 12. The secondary efficacy outcome measures forthis study were as follows: (i) percent and absolute change frombaseline in EASI score at Week 12; (ii) percent of patients achieving a75% reduction from baseline in EASI score (EASI-75) at Week 12; (iii)percent of patients achieving an IGA score of 0 or 1 at Week 12; (iv)percent of patients with a 2 point reduction from baseline in IGA atWeek 12; (v) absolute change from baseline in IGA at Week 12; (vi)percent of patients achieving an IGSA score of 0 or 1 at Week 12; (vii)percent of patients with a 2 point reduction from baseline in IGSA atWeek 12; (viii) absolute change from baseline in IGSA at Week 12; (ix)percent and absolute change from baseline in SCORAD at Week 12; (x)percent of patients with a 50% or 75% reduction from baseline inSCORAD-50/75 at Week 12; (xi) percent of patients achieving EASI-50 atWeek 12 and maintaining EASI-50 at Weeks 16 and 20; (xii) percent ofpatients achieving IGA score of 0 or 1 at Week 12 and maintaining IGAscore of 0 or 1 at Weeks 16 and 20; (xiii) percent of patients achievingIGSA score of 0 or 1 at Week 12 and maintaining IGSA score of 0 or 1 atWeeks 16 and 20; (xiv) percent of patients achieving SCORAD-50 at Week12 and maintaining SCORAD-50 at Weeks 16 and 20; (xv) percent changefrom baseline in total % body surface area (BSA) affected at Week 12;(xvi) absolute- and percent-change from baseline in pruritus as measuredby the Pruritus VAS (assessed as part of the SCORAD) at Week 12; (xvii)absolute- and percent-change from baseline in pruritus as measured bythe 5-D Itch Scale at Week 12; (xviii) total use (grams) of TCS frombaseline to Week 12; (xvix) total use (grams) of TCS from Week 12 to endof study or early termination; (xx) number of disease flares frombaseline to Week 12; (xxi) change in AD symptoms from baseline to Week12, as assessed by the ADSD; (xxii) change in AD-specific health-relatedQoL from baseline to Week 12, as assessed by the ADIQ; and (xxiii)change in health-related QoL from baseline to Week 12, as measured bythe DLQI.

Safety Outcome Measures. The safety outcome measures for this study wereas follows: (i) frequency and severity of treatment-emergent adverseevents; (ii) incidence of human anti-therapeutic antibodies (ATA) atbaseline and during the study; (iii) frequency and severity of skin andother organ system infections throughout the study; the clinicaldefinition of skin infection was as follows: the diagnosis of infectionwas based on the Investigator's clinical assessment including but notlimited to the presence of honey-colored crusting, serous discharge,pustules, or pain at the site of rash and that may be associated withsystemic features, including fever or a flare in AD disease (definedabove); and (iv) incidence of disease rebound following discontinuationof study drug as assessed by the investigator; the clinical definitionof disease rebound was: a significant worsening of disease severityafter cessation of therapy to a severity level that is greater thanprior to commencing therapy (Hijnen et al., J Eur Acad Dermatol Venereol2007; 21(1) 85-9).

Atopic Dermatitis Impact Questionnaire (ADIQ). We developed anAD-specific health-related quality of life tool for use in patients aged12 years and older following U.S. Food and Drug Administrationpatient-reported outcome (PRO) guidance (available online atwww(dot)fda(dot)gov(slash)downloads(slash)Drugs(slash)Guidances(slash)UCM193282(dot)pdf. Specifically, a literature review was conducted to assess the ADPRO landscape. Concept elicitation (CE) interviews with adults ages18-75 (n=15) and adolescents 12-17 (n=15) with moderate or severe ADwere conducted to elicit signs and symptoms of AD that are important topatients. A grounded theory approach was used to qualitatively analyzeinterview transcripts. An item generation meeting (IGM) was held toreview the data and develop the preliminary ADIQ based on patientfeedback from the CE interviews.

The draft ADIQ was tested in cognitive interviews with 34 adult andadolescent patients who confirmed clarity and relevance of the conceptsincluded in the ADIQ, shown in Table 2 below. The psychometricvalidation is not included.

TABLE 2 Atopic Dermatitis Impact Questionnaire (ADIQ). Please select theresponse that best describes how you have been during the past 7 dayswith regards to your atopic dermatitis. Question Response Choices 1.Over the last 7 days, how depressed were 0 = Not at all depressed youbecause of your atopic dermatitis? 1 = A little depressed 2 = Somewhatdepressed 3 = Quite a bit depressed 4 = Extremely depressed 2. Over thelast 7 days, how anxious were 0 = Not at all anxious you because of youratopic dermatitis? 1 = A little anxious 2 = Somewhat anxious 3 = Quite abit anxious 4 = Extremely anxious 3. Over the last 7 days, how angrywere you 0 = Not at all angry because of your atopic dermatitis? 1 = Alittle angry 2 = Somewhat angry 3 = Quite a bit angry 4 = Extremelyangry 4. Over the last 7 days, how frustrated were 0 = Not at allfrustrated you because of your atopic dermatitis 1 = A little frustrated2 = Somewhat frustrated 3 = Quite a bit frustrated 4 = Extremelyfrustrated 5. Over the last 7 days, how embarrassed 0 = Not at allembarrassed were you when you were around other people 1 = A littleembarrassed because of your atopic dermatitis? 2 = Somewhat embarrassed3 = Quite a bit embarrassed 4 = Extremely embarrassed 6. Over the last 7days, how self-conscious 0 = Not at all self-conscious were you becauseof your atopic dermatitis? 1 = A little self-conscious 2 = Somewhatself-conscious 3 = Quite a bit self-conscious 4 = Extremelyself-conscious 7. Over the last 7 days, how much has your 0 = Not at allatopic dermatitis affected your self-esteem? 1 = A little 2 = Somewhat 3= Quite a bit 4 = A lot 8. Over the last 7 days, how much has your 0 =Not at all atopic dermatitis interfered with your sex 1 = A little life?2 = Somewhat 3 = Quite a bit 4 = A lot 9. Over the last 7 days, howtired did you 0 = Not at all tired feel because of your atopicdermatitis? 1 = A little tired 2 = Somewhat tired 3 = Quite a bit tired4 = Extremely tired 10. Over the last 7 days, how much 0 = Nonedifficulty did you have falling asleep because 1 = A little of youratopic dermatitis? 2 = Some 3 = Quite a bit 4 = A lot 11. Over the last7 days, how much did your 0 = None atopic dermatitis interfere with yourability to 1 = A little work at your job or at school? 2 = Some 3 =Quite a bit 4 = A lot 12. Over the last 7 days, how often did you 0 =None of the time miss work or school because of your atopic 1 = A littleof the time dermatitis? 2 = Some of the time 3 = Most of the time 4 =All of the time 13. Over the last 7 days, how often did you 0 = None ofthe time stay at home because of your atopic 1 = A little of the timedermatitis? 2 = Some of the time 3 = Most of the time 4 = All of thetime 14. Over the last 7 days, how often did you 0 = None of the timeavoid crowded placed because of your atopic 1 = A little of the timedermatitis? 2 = Some of the time 3 = Most of the time 4 = All of thetime 15. Over the last 7 days, how much did your 0 = Not at all atopicdermatitis interfere with your 1 = A little relationships with friends?2 = Somewhat 3 = Quite a bit 4 = A lot 16. Over the last 7 days, howmuch did your 0 = Not at all atopic dermatitis interfere with yoursocial 1 = A little activities? 2 = Somewhat 3 = Quite a bit 4 = A lot17. Over the last 7 days, how often were you 0 = None of the time unableto wear the clothes you wanted to 1 = A little of the time wear becauseof your atopic dermatitis? 2 = Some of the time 3 = Most of the time 4 =All of the time

Results

In this study, 209 patients were treated. At Week 12, a greaterproportion of patients treated with lebrikizumab (125 mg every fourweeks) plus TCS achieved EASI-50 (FIG. 2A and FIG. 3A), EASI-75 (FIG.3B), EASI-90 (FIG. 3C) and SCORAD-50 (FIG. 2B) compared with placebo. Inaddition, At Week 12, the percentage of patients who achieved EASI-50,the primary efficacy outcome, was greater for all lebrikizumab groupscompared with placebo (62.3%), but only achieved statisticalsignificance in the multiple-dose arm (82.4% [p=0.026] for the 125 mgQ4W arm) as shown in FIG. 11A. Overall, the proportion of patientsachieving an EASI-75 response, a secondary efficacy outcome, was greaterin all lebrikizumab dose groups, but was significantly greater only inthe 125 mg Q4W group (54.9% [p=0.036]) compared with placebo (34.0%) atWeek 12 as shown in FIG. 11B. The percentage of patients who achievedIGA 0/1 at Week 12 was higher in all lebrikizumab groups compared withplacebo, but did not reach statistical significance. However, as withthe other outcomes, the data suggest a dose—response relationship forIGA (FIG. 11C), and the 125 mg Q4W group showed continued improvementsin the final weeks of the treatment period (FIG. 2C). With regards toSCORAD-50, more patients in the lebrikizumab 125 mg Q4W group (51.0%[p=0.018]) and 250 mg SD groups (47.2% [p=0.030]) achieved this endpointat Week 12 compared with placebo (26.4%; FIG. 11D). All lebrikizumabgroups showed improvements in BSA affected (Table 3). The greatestreduction in BSA affected at Week 12 was observed in the lebrikizumab125 mg Q4W group (57.7% reduction), although there were also substantiveimprovements in the placebo group (47.4%). Placebo-corrected efficacyfor BSA was not statistically significant (p=0.38).

Lebrikizumab treatment led to improvements in Pruritus VAS (Table 3 andTable 7). Improvements in Pruritus VAS during the 2-week TCS run-inperiod were quantitatively larger than improvements in severity measureoutcomes. There was a high compliance rate of TCS use amongst alltreatment groups with TCS used 86.8% (125 mg SD), 86.7% (250 mg SD),91.9% (125 mg Q4W) and 88.2% (placebo) of days on average from baselineto Week 12.

There were also greater improvements in sleep loss VAS, as well astrending improvement in IGA 0/1, pruritus VAS and AD specifichealth-related quality of life (QoL) measurement, specifically theAtopic Dermatitis Impact Questionnaire (ADIQ), in patients treated with125 mg lebrikizumab every 4 weeks plus TCS compared with placebo at Week12 (Table 3, Table 7, FIG. 12A, FIG. 12B, and FIG. 12C). There were alsodose-dependent improvements in DLQI scores, with numerically greaterreductions from baseline with all lebrikizumab groups compared withplacebo (Table 3, Table 7, and FIG. 12D).

Pharmacokinetic parameters were also assessed as follows. Serum samplesfor analysis of lebrikizumab pharmacokinetics were obtained at Day 1(pre-dose) and at Weeks 1, 4 (pre-dose), 6, 8 (pre-dose), 12, 16, and 20for all dosing regimens. Serum lebrikizumab concentrations weresummarized by treatment and visit using descriptive statistics for thepatients that received one of the lebrikizumab treatment regimens. Thereported PK parameters include the Week 1 C_(max), C_(min) at Weeks 4,8, and 12, and the elimination half-life. The mean PK parameters andrespective standard deviations for each of the lebrikizumab dosingregimens are shown in Table 8. The results show that thepharmacokinetics of lebrikizumab in AD patients were consistent with thepharmacokinetics for lebrikizumab reported in the published literaturepreviously for adult asthma (see, e.g., Hanania et al., Thorax 70(8):748-756 (2015)), showing linear and dose-proportional characteristicswith a half-life of 19-22 days.

Adverse event rates were generally similar between treatment groups(66.7% all lebrikizumab groups vs. 66.0% placebo) and most were mild ormoderate in severity (Table 9). Specifically, three (2%) patients in thelebrikizumab group (all doses combined) and one (2%) patient in theplacebo group experienced an AE (skin infection in the 125 mg SD group,anxiety and myopathy in 125 mg Q4W group and atopic dermatitis in theplacebo group) that led to withdrawal from the study. One (1%) patientin the lebrikizumab group experienced an AE (gastrointestinal viralinfection) that led to dose interruption. There were no deaths, eventsof anaphylaxis, malignancy, or protocol-defined parasitic, or targetedintracellular infections of interest. Injection-site reactions occurredinfrequently (1.3% all lebrikizumab groups and 1.9% placebo); all eventswere non-serious, lasted a median of 1-3 days, and did not lead totreatment discontinuation.

Herpes infections occurred infrequently and only amonglebrikizumab-treated patients (n=6 [3.8%]; herpes simplex in n=4 [2.6%]and herpes zoster in n=2 [1.3%]); all events were non-serious, and nonelead to treatment discontinuation. In addition, there were no events ofeczema herpeticum. There were 14 (9%) patients with skin infectionsamong the lebrikizumab-treated groups (all doses combined) and nine(17%) patients in the placebo group. Eosinophil-associated AEs were alsoreported infrequently and only occurred among five lebrikizumab-treatedpatients (3.2%; 3 events of “eosinophilia” and 2 events of “eosinophilcount increased”); all events were non-serious, mild to moderate inintensity, and did not result in interruption of treatment. The maximumeosinophil count in these five patients ranged from 0.91 to 3.2×10⁹/Land of these, three were Grade 2 eosinophilia (1501-5000 cell/mm³). Ofthese five patients, there were no associated clinical symptoms notedwith the reported events. Furthermore, the increases observed were inline with what has been seen in previous lebrikizumab studies (Hananiaet al., Thorax 2015:70(8):748-756; Hanania et al., Lancet Respir Med2016; 4(10): 781-796). We evaluated conjunctivitis given previousimbalances reported in biologic trials in this disease area (Thaci etal., Lancet 2016: 387 (10013): 40-52). A total of 15 (9.6%) patients inthe pooled lebrikizumab group and four patients (7.5%) in the placebogroup had a conjunctivitis AE; all events were non-serious and none ledto treatment discontinuation.

TABLE 3 Key efficacy results at Week 12. Lebri 250 mg Lebri 125 mg Lebri125 mg Single Dose + Single Dose + Q4W + TCS Placebo + TCS TCS BID TCSBID BID BID Endpoint (n = 53) (n = 52) (n = 51) (n = 53) % Patientsachieving EASI-50 N (%) 37 (69.8) 36 (69.2) 42 (82.4) 33 (62.3) Placebo-7.6 7.0 20.1 corrected changes p-value 0.44 0.48 0.026 % Patientsachieving EASI-75 N (%) 26 (49.1) 20 (38.5) 28 (54.9) 18 (34.0) Placebo-15.1 4.5 20.9 corrected changes p-value 0.12 0.66 0.036 % Patientsachieving SCORAD-50 N (%) 25 (47.2) 18 (34.6) 26 (51.0) 14 (26.4)Placebo- 20.8 8.2 24.6 corrected change p-value 0.030 0.38 0.012 SCORADscore % change from baseline N 53 52 51 53 Adjusted mean −42.6 (4.07)−38.7 (4.14) −53.5 (4.22) −35.4 (4.16) (SE) Δ (95% CI) −7.2 (−18.7, 4.3)−3.3 (−14.9, 8.3) −18.0 (−29.7, −6.4) p-value 0.22 0.57 0.0026 %Patients achieving SCORAD-75 N (%) 6 (11.3) 7 (13.5) 11 (21.6) 7 (13.2)% Patients achieving IGA 0/1 N (%) 15 (28.3) 11 (21.2) 17 (33.3) 10(18.9) Placebo- 9.4 2.3 14.5 corrected change p-value 0.26 0.77 0.098Total % BSA affected, % change from baseline N 53 52 51 53 Adjusted mean−38.6 (8.07) −45.2 (8.21) −57.7 (8.35) −47.4 (8.24) (SE) Δ (95% CI) 8.8(−13.9, 31.6) 2.2 (−20.7, 25.1) −10.3 (−33.4, 12.9) p-value 0.45 0.850.38 % change from baseline in Prurit us VAS (from SCORAD) Mean −32.82−34.92 −40.71 −27.54 Placebo- −5.28 −7.38 −13.7 corrected change p-value0.54 0.40 0.13 % change from baseline in Sleep Loss VAS (from SCORAD)Mean (SD) −47.2 −53.1 −53.6 −22.6 Placebo- −24.6 −30.6 −31.0 correctedchange p-value 0.08 0.03 0.02 % change from baseline in AD Specifichealth-related QoL - ADIQ Mean −30.8 −33.9 −54.3 −29.0 Placebo- −1.8−4.9 −25.3 corrected change p-value 0.89 0.71 0.052 DLQI, % change frombaseline N 53 52 51 53 Adjusted mean −40.67 (6.69) −34.33 (6.93) −43.12(7.02) −33.57 (6.93) (SE) Δ (95% CI) −7.09 (−26.04, 11.85) −0.76(−20.04, 18.52) −9.55 (−28.91, 9.82) p-value 0.46 0.94 0.33 ADIQ =Atopic Dermatitis Impact Questionnaire; BSA = body surface area; CI =confidence interval; DLQI = Dermatology Life Quality Index; EASI =Eczema Area Severity Index; IGA = Investigator Global Assessment; SCORAD= SCORing Atopic Dermatitis; SE = standard error; VAS = Visual AnalogScale.

TABLE 7 Percent change in EASI, SCORAD, IGA, Pruritus VAS, and percentBSA affected from screening to baseline (run-in). LebrikizumabLebrikizumab Lebrikizumab 125 mg SD 250 mg SD 125 mg Q4W Placebo Allpatients (n = 52) (n = 53) (n = 51) (n = 53) (n = 209) Mean % Change in−14.3 (27.6) −10.2 (29.2) −9.1 (22.7) −6.4 (32.2) −10.0 (28.1) EASI (SD)Mean Absolute −0.21 (0.50) −0.17 (0.51) −0.14 (0.40) −0.09 (0.45) −0.15(0.47) Change in IGA (SD) Mean % Change in −12.2 (20.0) −9.9 (14.7) −7.7(15.0) −5.0 (20.3) −8.7 (17.8) SCORAD (SD) Mean % Change in −17.1 (26.7)−4.3 (26.9) −9.3 (27.0) −6.2 (23.2) −9.2 (26.3) % BSA Affected (SD) Mean% Change in −23.8 (29.6) −15.5 (28.6) −15.2 (39.4) −12.8 (29.8) −16.8(32.1) Pruritus VAS (SD) Mean % Change in −29.0 (43.6) −8.1 (81.8) −12.9(40.1) −5.4 (57.2) −13.9 (58.2) Sleep VAS (SD) BSA = body surface area;EASI = Eczema Area Severity Index; IGA = Investigator Global Assessment;Q4W = every 4 weeks; SD = single dose SCORAD = SCORing AtopicDermatitis; SD = single dose; SE = standard error; VAS = Visual AnalogScale.

TABLE 8 Mean Lebrikizumab Pharmacokinetic Parameters Following Single orMultiple Dose Subcutaneous Administration. Lebrikizumab LebrikizumabLebrikizumab 125 mg SD 250 mg SD 125 mg Q4W Mean C_(max, wk 1) 17.0(5.22) 35.6 (10.8) 16.1 (5.19) μg/mL (SD) Mean C_(min, wk 4) 10.2 (3.00)21.4 (6.87) 9.15 (2.99) μg/mL (SD) Mean C_(min, wk 8) 4.59 (2.12) 9.53(3.53) 13.6 (5.34) μg/mL (SD) Mean C_(min wk 12) 2.28 (1.72) 3.77 (2.01)14.4 (5.69) μg/mL (SD) t½, day 18.5 (5.06) 22.2 (6.18) 20.9 (4.17)C_(max, wk 1) = maximum lebrikizumab concentration at Week 1,C_(min, wk 4) = observed minimum concentration at Week 4, C_(min, wk 8)= observed minimum concentration at Week 8, C_(min, wk 12) = observedminimum concentration at Week 12, t_(1/2) = elimination half-life

TABLE 9 Overview of key safety information, Weeks 0 to 20. LebrikizumabLebrikizumab Lebrikizumab All 125 mg SD 250 mg SD 125 mg Q4Wlebrikizumab Placebo (n = 54) (n = 52) (n = 50) (n = 156) (n = 53)Patients with at least one 38 (70) 39 (75) 27 (54) 104 (67) 35 (66) AE,n (%) AE leading to withdrawal 1 (2) 0 2 (4) 3 (2) 1 (2) from study drugAE leading to dose 0 1 (2) 0 1 (1) 0 modification/interruption Patientswith at least one 3 (6) 0 2 (4) 5 (3) 2 (4) serious AE, n (%) SAEleading to withdrawal 0 0 1 (2) 1 (1) 0 from study drug Myopathy Adverseevents of interest, n (%) Adjudicated anaphylaxis 0 0 0 0 0 perSampson's criteria* Infections 24 (44) 20 (39) 12 (24) 56 (36) 24 (45)Patients with skin 6 (11) 5 (10) 3 (6) 14 (9) 9 (17) infection Injectionsite reactions 0 0 2 (4) 2 (1) 1 (2) Malignancies 0 0 0 0 0 Herpesinfections, n (%) Total number of patients 2 (3.9) 3 (5.7) 2 (3.9) 6(3.8) 0 with at least one infection Total number of infections 0 0 0 0 0related to study drug^(†) Herpes Simplex 1 (2.0) 2 (3.8) 1 (2.0) 4 (2.6)0 Herpes Zoster 1 (2.0) 1 (1.9) 1 (2.0) 2 (1.3) 0 Conjunctivalinfections, irritations and inflammations, n (%) Total number ofpatients 4 (7.4) 3 (5.8) 2 (4.0) 9 (5.8) 0 with at least one AE Totalnumber of events 4 (7.4) 3 3 10 0 Conjunctivitis allergic 4 (7.4) (3.8)2 (4.0) 8 (5.1) 0 Conjunctival hyperaemia 0 1 (1.9) 0 1 (0.6) 0 AE =adverse event; Q4W = every 4 weeks; SAE = serious adverse event; SD =single dose. *Blinded data was reviewed to adjudicate cases asanaphylaxis per Sampson's criteria. ^(†)lnfections related to study drugwere investigator-assessed.

We also observed an apparent dose-response relationship for the datashown in FIG. 2A. The 250 mg single dose was numerically better at earlytime points compared to the 125 mg single dose. Furthermore, the datashown in FIG. 2A suggests that the 125 mg dose once every four weeks didnot appear to have reached an efficacy plateau at Week 12. These datasuggest the potential benefit of either higher dosing (e.g., 250 mg Q4W)or a loading dose.

In addition, at Week 20, in the modified intention-to-treat population,a greater proportion of patients treated with 125 mg lebrikizumab every4 weeks plus TCS achieved EASI-50 (FIG. 3A), EASI-75 (FIG. 3B), andEASI-90 (FIG. 3C) compared with placebo. See also Table 9. A greaterpercent decrease from baseline in EASI over 12 weeks in thelebrikizumab-treated patients compared with placebo was also observed(FIG. 4). Notably, the patients who received 125 mg lebrikizumab onceevery four weeks for 12 weeks showed a 70.5% change (decrease) in EASIcompared to a 53.1% change (decrease) in the placebo arm. That 17.4%difference between lebrikizumab treatment and placebo was statisticallysignificant, P=0.025.

TABLE 9 Summary of key efficacy outcomes at Week 20. LebrikizumabLebrikizumab Lebrikizumab 125 mg SD 250 mg SD 125 mg Q4W PlaceboPatients maintaining EASI-50 response at Weeks 12 and 20 N (%) 29 (80.6)29 (78.4) 38 (90.5) 24 (72.7) Placebo-corrected differences (SE) 7.835.65 17.75 p-value 0.39 0.58 0.047 Patients maintaining EASI-75 responseat Weeks 12 and 20 N (%) 9 (45.0) 16 (61.5) 21 (75.0) 12 (66.7)Placebo-corrected differences (SE) −21.70 −5.13 8.33 p-value 0.202 0.810.54 Patients maintaining IGA 0/1 response at Weeks 12 and 20 N (%) 6(54.5) 9 (60.0) 12 (70.6) 1.6 (60) Placebo-corrected differences (SE)−5.45 0 10.59 p-value 0.97 0.94 0.58 Patients maintaining SCORAD-50response at Weeks 12 and 20 N (%) 11 (61.1) 16 (64.0) 13 (50.0) 11(78.6) Placebo-corrected differences (SE) −17.50 −14.60 −28.60 p-value0.34 0.49 0.13 Change from baseline in EASI Mean % change (SE) −62.1(5.4) −55.9 (5.3) −71.1 (5.5) −54.1 (5.5) Placebo-corrected differences(SE) −8.08 (7.72) −1.88 (7.62) −17.08 (7.78) p-value 0.30 0.81 0.030Change from baseline in the % BSA affected Mean % change (SE) 53.7 (6.1)−46.4 (6.0) 63.8 (6.2) 52 (6.2) Placebo-corrected differences (SE) 5.56(8.6) p-value 0.85 6.52 0.18 Change from baseline Pruritus VAS Mean %change (SE) −27.59 −30.25 −35.23 −21.46 Placebo-corrected differences(SE) −6.13 −8.79 −13.76 p-value 0.57 0.41 0.21 BSA = body surface area;EASI = Eczema Area Severity Index; IGA = Investigator Global Assessment;SCORAD = SCORing Atopic Dermatitis; SD = single dose; SE = standarderror; VAS = Visual Analog Scale.

We also measured patients' symptoms and health-related quality of lifeby SCORAD Pruritus Visual Analog Scale (VAS), Sleep Loss VAS and theADIQ. Improvements from Baseline were demonstrated in the Pruritus VAS,Sleep Loss VAS, and ADIQ (shown in Table 3). Post hoc analysis alsoexamined change from screening. Patients treated with 125 mglebrikizumab administered once every four weeks showed a Pruritus VAS55.2% (p=0.03 vs placebo) reduction from screening at Week 12 whileplacebo resulted in reductions of 39.3% from screening. There were alsoimprovements in Sleep Loss VAS from screening to Week 12, with a 61.5%(p=0.04 vs placebo) reduction with lebrikizumab treatment and placeboreduction 39.3%. There was a 64.7% (p=0.02 vs placebo) reduction in ADIQfrom screening with lebrikizumab treatment and placebo reduction of41.8%. Differences in these endpoints, relative to placebo, for groupsin which lebrikizumab was only given as a single dose (125 mg and 250mg) were in most cases not statistically significant.

In summary, these results demonstrate that targeting IL-13 inmoderate-to-severe AD provides clinically meaningful placebo-correctedimprovements in a number of severity outcomes, generally in adose-dependent manner. Monthly dosing significantly improved theproportion of patients achieving EASI-50, EASI-75, and SCORAD-50 withtrends for improvement in patients achieving IGA 0/1 and improvements inpruritus VAS. These improvements were seen on top of intensive TCSapplication associated with substantial responses in the placebo group.Adverse event rates were generally similar between treatment groups andmost were mild or moderate in severity.

In addition, the results show that lebrikizumab provides treatmentbenefit on top of rigorous application of TCS in patients withmoderate-to-severe AD who had an inadequate response to standard-of-careTCS treatment and baseline characteristics indicating a patientpopulation towards the more severe end of the spectrum. The study metits primary endpoint—the percentage of patients with EASI-50 at Week12—with a statistically significant number of patients in thelebrikizumab 125 mg Q4W group achieving this level of reduction in thebaseline EASI score. Moreover, the upward sloping response curves overthe final weeks of the treatment period suggest that the responseplateau might not have been reached by Week 12 for the lebrikizumab 125mg Q4W group and longer treatment duration may lead to improvedefficacy. There also appeared to be a dose—response relationship, withthe lebrikizumab 125 mg Q4W dose showing numerically highest responserates across most endpoints, in particular EASI, with lebrikizumab 125mg SD showing the least benefit at Week 12.

Notably, the lebrikizumab 250 mg SD group showed numerically higherresponses at earlier time points for several outcomes, suggesting thepotential benefit of either higher dosing or a loading dose. In all mainefficacy endpoints, there was a continuous improvement observed over thecourse of the 12-week treatment period. Differences in many of thesecondary efficacy endpoints at Week 12 were statistically significantat the unadjusted 5% significance level for patients in the lebrikizumab125 mg Q4W group compared with patients in the placebo group. Theseincluded the percentage of patients with EASI-75, percentage of patientswith SCORAD-50, percent change from baseline in EASI score, and percentchange from baseline in SCORAD. Improvements in IGA and Pruritus VASwere numerically higher than placebo. Pruritus is considered a keycontributor to the reduced QoL of patients with moderate-to-severe AD,and there were numerically greater improvements in QoL (as measured byDLQI and ADIQ) and sleep with the lebrikizumab 125 mg Q4W dosing, likelydue to improvements in pruritus. In general, the effects of lebrikizumabwere also maintained during the 8-week follow-up period in severaloutcomes, including EASI, IGA (0/1) and SCORAD responses maintaineduntil Week 20.

Given the dose-response relationships observed across multipleendpoints, and the trends towards improved efficacy with increasing doseand duration, it is reasonable to hypothesize that further increases inthe dose (either in the form of a loading dose or a higher dose such as250 mg Q4W) and/or treatment duration may lead to improved efficacy.Furthermore, the observed differences in the lebrikizumab dose-responserelationship between FEV₁ in asthma patients (Hanania et al., Thorax2015;70(8):748-756) and EASI/IGA endpoints in AD patients suggest thatAD may in fact require higher doses of lebrikizumab to achieve aresponse plateau. This would be consistent with a higher IL-13 burdenamong atopic dermatitis patients relative to asthma patients.

The protocol for this study required patient compliance with twice-dailyTCS during the 2-week run-in period to be eligible for randomization.Moreover, patients were only eligible for randomization if theymanifested sufficient AD severity after this run-in period. Although thepatients included in this study were being inadequately controlled byTCS, inclusion of this TCS run-in period led to substantial improvementof the disease. This improvement was demonstrated by the suppressedbaseline AD severity scores, especially itch measures, likely causingdemonstration of additional improvement beyond baseline to be moredifficult.

During the treatment period, the protocol mandated continued applicationof twice-daily TCS and patients were provided daily reminders byelectronic diaries; patients in this study applied TCS with a highcompliance rate of 88% in all treatment groups. While daily TCS use istypically recommended for acute lesions rather than continuously andindefinitely, this proof-of-concept Phase 2 study sought to understandthe potential efficacy of lebrikizumab in addition to intensely rigorousTCS application (Eichenfield et al., J Am Acad Dermatol.2014;71(1):116-132) and not to assess TCS sparing. Consideration wasgiven to less frequent application of TCS because this TCS regimen maynot be reflective of long-term treatment or of clinical practice.However, this regimen is consistent with TCS labeling, and there wereregulatory concerns about off-label usage at a lower frequency thanmandated by product labeling. At the same time, there were concerns thatan alternate design that did not allow for background TCS, such aslebrikizumab monotherapy, would lead to substantial patient dropoutand/or imputed patient failure within the control arm. Indeed, studiesof biologic therapies in a monotherapy setting have led todrop-out/imputed patient failure rates on the order of approximately 50%within the control arm (Simpson et al., N Engl J Med.2016;375(24):2335-2348) in contrast to the dropout rate of 13% for theplacebo arm in this study. The rigorous TCS application during the12-week treatment period likely explains the substantial responseobserved in the placebo arm. Prolonged and frequent TCS use has beenshown to result in progressive improvements in AD, but most guidelinessuggest limiting daily use to avoid AEs. See, e.g., Brunner et al., JAllergy Clin Immunol. 2016;138(1):169-178; Schneider et al., J AllergyClin Immunol. 2013;131(2):295-9.e1-27; and Hanifin et al., J Am AcadDermatol. 2004;50(3):391-404. Furthermore, this substantial response inthe placebo arm could be why a larger treatment difference between theplacebo and the lebrikizumab treatment arms was not observed. Despitethe relatively high efficacy of prolonged and frequent TCS use, therewere still significant improvements, particularly in AD signs (EAST) andglobal scores (SCORAD), with adding lebrikizumab treatment.

Dupilumab, an anti-IL-4Rα monoclonal antibody, demonstrated efficacy inpatients with moderate-to-severe AD and has been recently approved inAD. IL-4Rα is thought to be an important receptor subunit for both IL-4and IL-13 signaling. Studies of dupilumab in AD patients provide insightinto the potential of IL-13 blockade to treat AD, with the caveat thatthe relative importance of IL-4 compared with IL-13 in AD has not beenestablished. Both IL-13 and IL-4 share overlapping biology and effectorfunctions; for example, both play a role in T-helper 2-cell development,IgE production, eosinophil recruitment, epithelial barrier integritydisruption via down-regulation of antimicrobial peptides, fibrosis, anddecreased filaggrin expression. See, e.g., Paternoster et al., NatGenet. 2011;44(2):187-192; Kagami et al., Clin Exp Immunol.2005;141(3):459-466. Because of such high overlap in biology, blockadeof IL-13 alone could potentially provide comparable improvements in ADto blockade of IL-13 and IL-4 in combination, with a more specifictargeted action. In addition, targeting a soluble cytokine, such asIL-13 may offer the advantage of alinear PK profile with resultingimprovements in dosing frequency. This in part explains the ability todose lebrikizumab Q4W and may allow for less frequent dosing duringmaintenance. In contract, receptor targeting is associated withtarget-mediated drug clearance that may lead to rapid declines inconcentration after drug discontinuation or interruption, as is the casewith dupilumab, which is dosed Q2W (Kovalenko et al., CPTPharmacometrics Syst Pharmacol. (2016) Nov;5(11):617-624).

The results of this study suggest that IL-13-mediated signaling pathwaysplay an important role in the pathogenesis of AD, and the blockade ofthis cytokine could lead to significant clinical benefit. Patients withmoderate-to-severe AD showed improvements with lebrikizumab treatment,even with single doses and intensive TCS use. Future studies of longerduration, with more frequent or higher dosing and in a largerpopulation, on different background regimens (or absence of) will helpclarify the role of targeting IL-13 alone in AD.

Example 3 Alternative Dosing Regimens

To evaluate whether alternative lebrikizumab dosing regimens could bedeveloped and could be predicted to demonstrate efficacy, we developed anovel, mechanism-based, longitudinal PK-PD model as described below. Themodel was used to simulate the predicted efficacy, e.g., EASI-75 score,of various alternative dosing regimens, for example, including but notlimited to, various loading doses followed subsequently by variousmaintenance doses, e.g., flat doses of lebrikizumab administered atregular time intervals, or various lower flat doses or higher flatdoses, or increased treatment intervals with lebrikizumab.

The model development began with raw pharmacokinetic and efficacy datathat had been collected during the conduct of the Phase II ClinicalStudy I described above. The lebrikizumab pharmacokinetic data in atopicdermatitis patients was consistent with predictions made from thelebrikizumab population PK model, which had been developed on the basisof drug concentrations obtained from asthma patients and healthyvolunteers (data not shown). This allowed for the use of thelebrikizumab population PK model to facilitate the development of theatopic dermatitis PK-PD model as described below.

The atopic dermatitis PK-PD model of lebrikizumab was developed based onEASI scores up to 12 weeks from Clinical Study I and the previouslyestablished lebrikizumab population PK model. The PD response effect wasthe reduction in EASI score. The effects of placebo and/or TCSadministration were also incorporated into the model. Covariate analysissuggested that baseline EASI score was a significant covariate on bothK_(out), which is the tissue repair rate constant, and E_(con), which isthe placebo/TCS effect. The final model is shown in FIG. 5 and the modelparameters are provided in Table 4.

TABLE 4 Lebrikizumab Atopic Dermatitis Model Parameters. parameterestimate 95% CI k_(out) (day⁻¹) 0.0540 (0.0379, 0.0819) E_(con) 0.268(0.200, 0.361) E_(max) 0.729 (0.363, 1.00)  EC₅₀ (μg/mL) 8.08 (0.612,44.1)  baseline effect on k_(out) −0.280 (−0.821, 1.42)  baseline effecton E_(con) −0.639 (−1.93, 0.235) ω_(k) _(out) 0.468 (0.171, 0.836) ω_(E)_(con) 1.05 (0.817, 1.16)  ρ_(k) _(out) × E_(con) −0.443 (−1.00, 0.204)σ 0.469 (0.427, 0.501) The confidence intervals are the 2.5^(th) and97.5^(th) percentiles of the parameters estimated for 1000 bootstrapsamples; see FIG. 5 drawing description for explanation of symbols.

The model includes certain assumptions, such as the assumption that theexposure-response relationship is similar across all drugconcentrations, however, we found that it adequately described thechange in raw EASI score observed in Study 1. Additionally, the modelwas able to predict EASI-50, EASI-75, and EASI-90 responses through Week12 from Study 1 very well despite not being designed for this purpose.This result provided additional validation for the model and increasedour confidence in its accuracy and robustness.

We used the final model to simulate a number of dosing regimens and thenwe predicted EASI scores at various times during treatment. The dosingregimens simulated are provided in Table 5 below. All simulations wereconducted over a 24-week time period. A dosing regimen of 125 mgadministered once every four weeks was used as a reference point tocompare all of the simulated results. As shown in Table 5, the simulateddosing regimens were broadly categorized into four groups: Group 1regimens were loading dose and maintenance dose permutations, Group 2regimens were 250 mg administered once every four weeks-basedpermutations, Group 3 regimens were 250 mg administered once every eightweeks-based permutations, and Group 4 regimens were lower dosingpermutations based on 37.5 mg administered once every four weeks.

TABLE 5 Dosing Regimens Simulated with the Lebrikizumab AD Model.Loading Dose Maintenance Dose Duration Group 250 mg at Day 1 125 mgevery four weeks 24 weeks 1 starting at Week 4 250 mg at Day 1, 250 125mg every four weeks 24 weeks 1 mg at Day 15 starting at Week 4 250 mg atDay 1 and 125 mg every four weeks 24 weeks 1 Day 29 starting at Week 8500 mg at Day 1 125 mg every four weeks 24 weeks 1 starting at Week 4None 250 mg every four weeks 24 weeks 2 starting at Day 1 500 mg at Day1 250 mg every four weeks 24 weeks 2 starting at Week 4 None 250 mgevery eight weeks 24 weeks 3 starting at Day 1 500 mg at Day 1 250 mgevery eight weeks 24 weeks 3 starting at Week 4 None 37.5 mg every fourweeks 24 weeks 4 125 mg at Day 1 37.5 mg every four weeks 24 weeks 4

Simulations of patients achieving EASI-75 over time with each of theGroup 1 lebrikizumab dosing regimens are shown in FIG. 6. The PK-PDmodel predicted that all loading dose permutations resulted in smallimprovements in EASI-75 response over the 125 mg once every four weeksregimen alone. This improvement was most evident in the middle portionof the curve (e.g. Day 56-84) and was mostly diluted out by later timepoints (Day 140-168).

Simulations of patients achieving EASI-75 over time with each of theGroup 2 lebrikizumab dosing regimens are shown in FIG. 7. The PK-PDmodel predicted that the 250 mg once every four weeks-based regimensbegan to show separation (i.e., an improvement in EASI-75) from the 125mg once every four weeks-regimen during the middle portion of the curve,and that separation was maintained at later time points.

Simulations of patients achieving EASI-75 over time with each of theGroup 3 lebrikizumab dosing regimens are shown in FIG. 8. The PK-PDmodel predicted that the 250 mg administered once every eight weeksregimens had comparable efficacy to the 125 mg administered once everyfour weeks regimen, with a slight improvement in off weeks (e.g., atWeek 12 and Week 20), and a slightly worse result during a Week beforethe next dose was given (e.g., at Week 8 and Week 16).

Simulations of patients achieving EASI-75 over time with each of theGroup 4 lebrikizumab dosing regimens are shown in FIG. 9. The PK-PDmodel predicted that the 37.5 mg administered once every four weeksregimens were less efficacious than the 125 mg administered once everyfour weeks dosing regimen.

In summary, the lebrikizumab atopic dermatitis PK-PD model predictedthat all “once every four weeks” lebrikizumab dosing regimens providedequivalent or better EASI-75 scores compared to EASI-75 scores achievedwith 125 mg administered once every four weeks (the dose clinicallytested in the Phase II Clinical Study I described above) with theexception of dosing regimens that included a dose of 37.5 mglebrikizumab administered once every four weeks. These simulationsshowed that 37.5 mg lebrikizumab administered once every four weeks waspredicted to be less efficacious, as assessed by EASI-75 scores, thanany of the other simulated dosing regimens.

The lebrikizumab atopic dermatitis PK-PD model also predicted that thebest EASI-75 result was obtained with 250 mg lebrikizumab administeredonce every four weeks (with or without a loading dose) for a treatmentduration of 16-24 weeks. The model also predicted that the inclusion ofa loading dose improved the EASI-75 result at earlier time points (e.g.,at 8 or 12 weeks), but that the impact of this improvement compared todosing regimens lacking a loading dose diminished over time (e.g., at 20or 24 weeks). Thus, inclusion of a loading dose may provide improvedtreatment benefit to patients early in the course of treatment. Finally,the model predicted that administration of 250 mg of lebrikizumab onceevery eight weeks resulted in EASI-75 improvements similar to theEASI-75 results predicted with administration of 125 mg of lebrikizumabonce every four weeks. The overall improvement in EASI-75 for the “250mg q8w regimen” was predicted to be somewhat higher during weeks whenthe 250 mg dose was not administered (e.g., at week 4) and slightlylower than the “125 mg q4w regimen” immediately prior to a 250 mg doseadministration (e.g., at week 8). Thus, the modeling results describedhere demonstrate that a number of lebrikizumab dosing regimens arepredicted to provide therapeutic benefit to atopic dermatitis patients.

Example 4 Clinical Study II

Clinical Study II was a Phase II, randomized, open label study toevaluate the safety and efficacy of lebrikizumab monotherapy in adultpatients (18-75 years of age) with persistent moderate to severe AD, whowere inadequately controlled by TCS. The Study II Schema is provided inFIG. 10.

Screening. Patients eligible to enroll in the Study had to meet alleligibility criteria described below.

The inclusion criteria included the following: age 18 to 75 years; ADdiagnosed by the Hanifin/Rajka criteria and that had been present for atleast 1 year at screening; moderate to severe AD as graded by theRajka/Langeland criteria at screening; history of inadequate response toa 1 month (within the 3 months prior to the screening visit) treatmentregimen of at least daily TCS and regular emollient for treatment of AD;EASI score 14 at screening and end of the run-in period (Visit 3), IGAscore 3 (5-point scale) at screening and end of the run-in period (Visit3); AD involvement of 10% body surface area (BSA) at screening; andPruritus VAS score (measured as part of the SCORAD) of 3 at screening;adherence to the protocol-specified 2-week run-in period TCS regimen (atleast 10 out of 14 days) at the time of entry into the treatment period(Day 1).

Exclusion criteria included the following: past and/or current use ofany anti-IL-13 or anti-IL-4/IL-13 therapy, including lebrikizumab; useof an investigational agent within 4 weeks prior to screening or within5 half-lives of the investigational agent, whichever is longer; historyof a severe allergic reaction or anaphylactic reaction to a biologicagent or known hypersensitivity to any component of the lebrikizumabinjection; hypersensitivity to TCS or to any other ingredients containedby the TCS product used in the study; use of any complementary,alternative, or homeopathic medicines including, but not limited to,phytotherapies, traditional or non-traditional herbal medications,essential fatty acids, or acupuncture within 7 days prior to the run-inperiod or need for such medications during the study; body weight <40 kgor body mass index >38 kg/m²; evidence of other skin conditions;including, but not limited to, T-cell lymphoma or allergic contactdermatitis; evidence of, or ongoing treatment (including topicalantibiotics) for active skin infection at screening (Day −15); certaininfections; history of recent or active (within 6 months) parasiticinfections, especially nematodes (e.g., Ascaris, Ancylostoma),Platyhelminthes (e.g., Schistosoma), or history of Listeria infections;active tuberculosis requiring treatment within the 12 months prior toVisit 1; evidence of acute or chronic hepatitis or known livercirrhosis; known immunodeficiency, including HIV infection; use of TCI(topical calcineurin inhibitor) at the time of screening; use of atanning booth/parlor within 4 weeks before the screening visit; allergenimmunotherapy within 3 months of screening; receipt of a live attenuatedvaccine within 4 weeks prior to baseline visit (Day 1); planned surgeryduring the study; clinically significant abnormality on screening ECG orlaboratory tests (hematology, serum chemistry, and urinalysis); AST,ALT, or total bilirubin elevation ≥2.0× the upper limit of normal (ULN)during screening; known current malignancy or current evaluation for apotential malignancy, including basal or squamous cell carcinoma of theskin or carcinoma in situ; history of malignancy within 5 years prior toscreening, except for appropriately treated carcinoma in situ of thecervix, non-melanoma skin carcinoma; stage I uterine cancer; and otherclinically significant medical disease that is uncontrolled despitetreatment.

Run-in Period. Following screening, eligible patients entered a 2-weekrun-in period (Days −14 to −1). After the run-in period, patientsstopped use of their pre-study TCS (and other AD medications) and begana protocol-specified topical therapy regimen as outlined below. On thefirst day of the run-in period (Day −14) patients received TCS for usethroughout the run-in period. Specifically, patients receivedtriamcinolone acetonide 0.1% cream for use on the body andhydrocortisone 2.5% cream for use on the face and intertriginous areas.Patients were instructed to apply emollient to all xerotic skin surfacesat least once daily and to apply cream to active skin lesions twice perday and not to apply cream to unaffected areas.

At the end of the run-in period, an assessment of disease severity wasperformed and patients who continued to demonstrate moderate to severeAD as assessed by EASI and IGA (see Inclusion Criteria above) wereeligible for randomization to study drug (lebrikizumab) or TCS alone.

Treatment Period (Weeks 1-12). At the end of the run-in period, patientswho had demonstrated adherence to the protocol-specified TCS regimen andcontinued to fulfill the eligibility criteria (see Inclusion Criteriaabove) were randomized. A total of approximately 50 patients wererandomized (1:1) to one of the following two treatment groups (see FIG.10): Group 1: 125 mg of lebrikizumab administered by subcutaneous (SC)injection every 4 weeks (Q4W) for a total of 3 doses; and Group 2: TCScream alone. Patients randomized to Group 1 did not receive aprotocol-specified TCS regimen and patients randomized to Group 2continued to apply TCS using the same TCS regimen that was appliedduring the run-in period (two times daily to lesional skin only).

Safety Follow-up Period (Weeks 13-20). All patients who completed studytreatment during the treatment period (Weeks 1-12) were followed forsafety for an additional 8 weeks (Weeks 13-20). During this safetyfollow-up period, patients in Group 2 (TCS only) were no longer requiredto apply the protocol specified TCS regimen as specified during therun-in and treatment periods. Instead, triamcinolone acetonide 0.1%cream and/or hydrocortisone 2.5% cream could be applied to active skinlesions only, as determined by the patient and the study investigator.Likewise, patients in treatment Group 1 (lebrikizumab monotherapy) coulduse triamcinolone acetonide 0.1% cream and/or hydrocortisone 2.5% creamon active skin lesions only, as determined by the patient and the studyinvestigator. All patients were encouraged to continue applyingemollients at least once daily to xerotic skin.

Topical calcineruin inhibitors (TCIs) were not allowed at any timeduring this study. Patients who had been using TCIs at the time ofscreening were allowed to participate in the study if they agreed tostop using TCIs during the study and if, in the opinion of theinvestigator, it was safe to stop the use of TCI and use the protocolspecified TCS cream instead.

Rationale for Lebrikizumab Dose and Schedule. The rationale for thelebrikizumab dose and schedule for Study II was similar to the rationaledescribed above for Study I. One lebrikizumab dose regimen (125 mg Q4W[Group 1]) was tested in this Phase II study. The 125-mg Q4W doseregimen was included as a potentially effective dose and was the highestdose regimen being studied in the pivotal Phase III adult studies(LAVOLTA I and II) of lebrikizumab in asthma. Given the expectedsimilarity in the role of IL-13 and the pharmacokinetics between asthmaand AD, it was hypothesized that this dose could be effective inpatients with AD.

As described above, however, the results of the LAVOLTA I and II studieswere inconsistent. Hanania et al., Lancet Respir Med 2016, available atdx(dot)doi(dot)org(slash)52213-2600(16)30265-X, published onlineSeptember 5, 2016. Both studies failed to show a clear dose-responsealthough pharmacokinetic and pharmacodynamics results were consistentwith those from the phase II studies described previously, indicatingthat drug exposures were similar and that the IL-13 pathway wasinhibited. Hanania et al., 2016, supra. Accordingly, it was uncertainwhether lebrikizumab would provide clinically meaningful benefit toatopic dermatitis patients at the dosing regimens described herein

The primary endpoint for the Phase II study was measured at Week 12. Themean serum lebrikizumab trough concentration at Week 12 was expected tobe over 90% of the steady state value for patients receiving 125 mg Q4W.In addition, for the 125 mg Q4W dose, the predicted troughconcentrations at Week 12 would be well within the range wherelebrikizumab demonstrated biological activity in the Phase II asthmaclinical trials.

Rationale for Control Group. Patients in treatment Group 2 received TCScream alone and therefore served as an active comparator. This treatmentgroup was used as a control for lebrikizumab SC to adequately assess thesafety and efficacy of lebrikizumab as monotherapy compared with TCS.

The active TCS comparator group was included since TCS use is thestandard of care in clinical practice (European Academy of Dermatologyand Venereology [EADV] 2009 guidelines; Darsow et al., J Eur AcadDermatol Venereol 2010; 24:317-28; American Academy of Dermatology 2014guidelines; Eichenfield et al., J Am Acad Dermatol 2014:71(1):116-32).

Efficacy Outcome Measures. The exploratory efficacy outcome measures oflebrikizumab monotherapy compared to TCS alone for this study were asfollows, in order: (i) percent of patients with a 50% or 75% (EAST50/75) reduction from baseline EASI at Week 12 (EASI-50 and EASI-75 weredefined as a 50% and 75% reduction, respectively, in EASI score at Week12 compared to baseline); (ii) percent and absolute change from baselinein EASI score at Week 12; (iii) percent of patients achieving an IGAscore of 0 or 1 at Week 12; (iv) percent of patients with a ≥2 pointreduction from baseline in IGA at Week 12; (v) absolute change frombaseline in IGA at Week 12; (vi) percent of patients achieving an IGSAscore of 0 or 1 at Week 12; (vii) percent of patients with a ≥2 pointreduction from baseline in IGSA at Week 12; (viii) absolute change frombaseline in IGSA at Week 12; (ix) percent and absolute change frombaseline in SCORAD at Week 12; (x) percent of patients with a 50% or 75%reduction (SCORAD 50/75) from baseline in SCORAD score at Week 12; (xi)percent change from baseline in total percent body surface area (BSA)affected at Week 12; (xii) absolute- and percent-change from baseline inpruritus as measured by the Pruritus VAS (assessed as part of theSCORAD) at Week 12; (xiii) number of disease flares from baseline toWeek 12; and (xiv) percent of patients who receive non-protocolspecified TCS before Week 12.

Safety Outcome Measures. The safety outcome measures for this study wereas follows: (i) incidence of treatment-emergent adverse events at Week12 with lebrikizumab used as monotherapy compared to TCS alone; (ii)incidence of human anti-therapeutic antibodies (ATA) at baseline andduring the study; (iii) frequency and severity of skin and other organsystem infections throughout the study; the clinical definition of skininfection was as follows: the diagnosis of infection was based on theInvestigator's clinical assessment including but not limited to thepresence of honey-colored crusting, serous discharge, pustules, or painat the site of rash and that may be associated with systemic features,including fever or a flare in AD disease (defined above); (iv) incidenceof disease rebound following discontinuation of study drug as assessedby the investigator; the clinical definition of disease rebound was: asignificant worsening of disease severity after cessation of therapy toa severity level that is greater than prior to commencing therapy(Hijnen et al., J Eur Acad Dermatol Venereol 2007; 21(1) 85-9); (v)incidence of injection site reactions from baseline to Week 12; and (vi)frequency and severity of treatment-emergent adverse events, includingadverse events of special interest from baseline to Week 12.

Results. The key efficacy results from Clinical Study II are summarizedin Table 6 below. The data show that lebrikizumab monotherapy over the12 week study period was effective. The EASI-75 and EASI-90 results weresimilar between the lebrikizumab and TCS treatment arms while the IGA,SCORAD, and pruritus results were somewhat lower in the lebrikizumab armthan in the TCS arm.

TABLE 6 Key Efficacy Results at Week 12. Baseline to Week 12 Day −14 toWeek 12 TCS lebrikizumab TCS lebrikizumab Endpoint (n = 27) (n = 28) (n= 27) (n = 28) EASI-75 37.0% 39.3% 37.0% 42.9% EASI-90 7.4% 14.3% 22.2%14.3% IGA 0/1 25.9% 7.1% 25.9% 7.1% % change in EASI −57.7% −44.0%−64.0% −53.8% % change in −41.3% −26.6% −48.9% −40.5% SCORAD % change inBSA −43.2% −43.2% −47.5% −52.2% % change in −34.9% −14.5% −53.2% −39.6%pruritus VAS* *in patients with baseline pruritus VAS ≥3.

Anti-11,13 Antibody (Lebrikizumab) Amino Acid Sequences

The table below shows the amino acid sequences of the CDR-H1, CDR-H2,CDR-H3, CDR-L1, CDR-L2, and CDR-L3 regions of lebrikizumab, along withVH, VL, heavy chain sequences and light chain sequences. As indicated inthe Table of Sequences below and as described above, VH and the heavychain may include an N-terminal glutamine and the heavy chain may alsoinclude a C-terminal lysine. As is well known in the art, N-terminalglutamine residues can form pyroglutamate and C-terminal lysine residuescan be clipped during manufacturing processes.

TABLE OF SEQUENCES SEQ ID NO: Description Sequence 1 lebrikizumabVTLRESGPAL VKPTQTLTLT CTVSGFSLSA YSVNWIRQPP heavy chainGKALEWLAMI WGDGKIVYNS ALKSRLTISK DTSKNQVVLT variable regionMTNMDPVDTA TYYCAGDGYY PYAMDNWGQG SLVTVSS 2 lebrikizumabDIVMTQSPDS LSVSLGERAT INCRASKSVD SYGNSFMHWY light chainQQKPGQPPKL LIYLASNLES GVPDRFSGSG SGTDFTLTIS variable regionSLQAEDVAVY YCQQNNEDPR TFGGGTKVEI K 3 AlternateQVTLRESGPA LVKPTQTLTL TCTVSGFSLS AYSVNWIRQP lebrikizumab VHPGKALEWLAM IWGDGKIVYN SALKSRLTIS KDTSKNQVVLTMTNMDPVDT ATYYCAGDGY YPYAMDNWGQ GSLVTVSS 4 AlternateDIVMTQSPDS LSVSLGERAT INCRASKSVD SYGNSFMHWY lebrikizumab VLQQKPGQPPKL LIYLASNLES GVPDRFSGSG SGTDFTLTISSLQAEDVAVY YCQQNNEDPR TFGGGTKVEI KR 5 lebrikizumab AYSVN HVR-H1 6lebrikizumab MIWGDGKIVYNSALKS HVR-H2 7 lebrikizumab DGYYPYAMDN HVR-H3 8lebrikizumab RASKSVDSYGNSFMH HVR-L1 9 lebrikizumab LASNLES HVR-L2 10lebrikizumab QQNNEDPRT HVR-L3 11 lebrikizumabQVTLRESGPA LVKPTQTLTL TCTVSGFSLS AYSVNWIRQP heavy chainPGKALEWLAM IWGDGKIVYN SALKSRLTIS KDTSKNQVVLTMTNMDPVDT ATYYCAGDGY YPYAMDNWGQ GSLVTVSSASTKGPSVFPLA PCSRSTSEST AALGCLVKDY FPEPVTVSWNSGALTSGVHT FPAVLQSSGL YSLSSVVTVP SSSLGTKTYTCNVDHKPSNT KVDKRVESKY GPPCPPCPAP EFLGGPSVFLFPPKPKDTLM ISRTPEVTCV VVDVSQEDPE VQFNWYVDGVEVHNAKTKPR EEQFNSTYRV VSVLTVLHQD WLNGKEYKCKVSNKGLPSSI EKTISKAKGQ PREPQVYTLP PSQEEMTKNQVSLTCLVKGF YPSDIAVEWE SNGQPENNYK TTPPVLDSDGSFFLYSRLTV DKSRWQEGNV FSCSVMHEAL HNHYTQKSLS LSLGK 12 lebrikizumabDIVMTQSPDS LSVSLGERAT INCRASKSVD SYGNSFMHWY light chainQQKPGQPPKL LIYLASNLES GVPDRFSGSG SGTDFTLTISSLQAEDVAVY YCQQNNEDPR TFGGGTKVEI KRTVAAPSVFIFPPSDEQLK SGTASVVCLL NNFYPREAKV QWKVDNALQSGNSQESVTEQ DSKDSTYSLS STLTLSKADY EKHKVYACEV THQGLSSPVT KSFNRGEC

1-63. (canceled)
 64. A method of treating atopic dermatitis in apatient, the method comprising administering to the patient an IL-13antibody, wherein administering comprises administering a loading doseof 250 mg or 500mg of the anti-IL-13 antibody and administering asubsequent maintenance dose of 125 mg of the anti-IL-13 antibody,wherein the loading dose is administered once or twice, and themaintenance dose is administered for the remainder of treatmentduration, wherein the anti-IL-13 antibody is an antibody comprising aheavy chain having the amino acid sequence of SEQ ID NO: 11 and a lightchain having the amino acid sequence of SEQ ID NO: 12, wherein theadministration of the anti-IL-13 antibody reduces disease severity inthe patient, and wherein the disease severity is assessed by an AtopicDermatitis Disease Severity Outcome Measure. 65-79. (canceled)
 80. Themethod of claim 64, wherein the maintenance dose is administered fourweeks after administration of the loading dose and wherein themaintenance dose is administered once every four weeks thereafter forthe duration of treatment. 81-83. (canceled)
 84. The method of claim 64,wherein the duration of treatment is 24 weeks or more.
 85. The method ofclaim 84, wherein the duration of treatment is 24 weeks.
 86. (canceled)87. The method of claim 64, wherein the Atopic Dermatitis DiseaseSeverity Outcome Measure is Eczema Area and Severity Index (EAST).88-89. (canceled)
 90. The method of claim 64, wherein the AtopicDermatitis Disease Severity Outcome Measure is Severity Scoring ofAtopic Dermatitis (SCORAD). 91-92. (canceled)
 93. The method of claim64, wherein the Atopic Dermatitis Disease Severity Outcome Measure isInvestigator Global Assessment (IGA). 94-95. (canceled)
 96. The methodof claim 64, wherein the Atopic Dermatitis Disease Severity OutcomeMeasure is a Patient Reported Outcome (PRO).
 97. The method of claim 96,wherein the PRO is pruritus visual analogue scale (VAS), sleep loss VAS,or Atopic Dermatitis Impact Questionnaire (ADIQ) score. 98-101.(canceled)
 102. The method of claim 64, wherein the atopic dermatitis ismoderate to severe as determined by Rajka/Langeland criteria score andwherein the Rajka/Langeland criteria score is determined to be between4.5 and
 9. 103. The method of claim 64, wherein the IL-13 antibody isadministered to the patient using a subcutaneous administration device.104. The method of claim 103, wherein the subcutaneous administrationdevice is selected from a prefilled syringe, disposable pen injectiondevice, microneedle device, microinfuser device, needle-free injectiondevice, and autoinjector device.
 105. The method of claim 64, whereinthe method further comprises administration of one or more topicalcorticosteroids.
 106. The method of claim 105, wherein the one or moretopical corticosteroids is or are administered before administration ofthe IL-13 antibody, at the same time as administration of the IL-13antibody, or after administration of the IL-13 antibody.
 107. The methodof claim 106, wherein the one or more topical corticosteroids is or areselected from triamcinolone acetonide, hydrocortisone, and a combinationof triamcinolone acetonide and hydrocortisone.
 108. The method of claim64, wherein the patient is 12 years of age or older.
 109. The method ofclaim 64, wherein the patient's atopic dermatitis is inadequatelycontrolled by topical corticosteroids.